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Daniel Fisch

@danielhfisch.bsky.social
324 followers 654 following 44 posts

Group Leader at the Gulbenkian Institute for Molecular Medicine (GIMM) Innate Immunology, Macrophages & Toll-Like Receptors 🔬👨‍🔬

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Daniel Fisch @danielhfisch.bsky.social · 02/10/2026
After Germany, the UK, France and the US, it feels wonderful to build my scientific home back in Europe. 🧪🇵🇹 Now the exciting part begins...building a team and chasing big questions together. Can't wait for the discoveries ahead!
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Daniel Fisch @danielhfisch.bsky.social · 02/10/2026
Of course I want to thank everyone who helped me along the way but especially my PhD and postdoc mentors @frickellab.bsky.social @avishenoy.bsky.social @jkagan1.bsky.social without whom the "Innate Immune Cell Biology Lab" at GIMM probably would not exist! ☺️
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Daniel Fisch @danielhfisch.bsky.social · 02/10/2026
Yesterday was a special day in my career: I got to open the door to my own research group (and for the first time in my life to my own office 😂)! Thank you to @gimminstitute.bsky.social for giving me this opportunity, it's exciting to finally be here!
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Daniel Fisch @danielhfisch.bsky.social · 24/09/2026
Thank you 😊
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Daniel Fisch @danielhfisch.bsky.social · 24/09/2026
Thanks Ivan ☺️
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Daniel Fisch @danielhfisch.bsky.social · 24/09/2026
Thank you 😊
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
@hfspo.bsky.social @embo.org @bostonchildrens.bsky.social
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
I want to thank all my colleagues from the Kagan lab for making this work possible and especially @jkagan1.bsky.social for his support of my postdoc training and mentorship for the past 5 years! Thanks to all funders supporting this work! (NIH, HFSP, EMBO, HRIA King Trust, CRI Irvington).
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
In summary: TRAM is identified as the release factor enabling receptor-free and sustained myddosome signaling in the TLR pathway. I hope you enjoy the read: rdcu.be/jGdKxErXyAsa
Graphical summary of the proposed model for TLR signaling.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
If long-lived myddosomes sustain inflammation, disrupting them should stop it. Conversely, MyD88 oligomerization inhibitors (MPP, TSI-13-57) dissolved already-formed myddosomes in cells. We detected for the first time native myddosomes in mice, which could be shut down by the same inhibitors.
Top: immunoblots of myddosome immunoprecipitations from splenocytes of mice injected with PBS, LTA or LPS and subsequently injected with TSI-13-57 or vehicle. IRAK2, IRAK4, TRAF6, IKKα, IKKβ, p65 and TBK1 co-precipitate with MyD88 after PAMP injection and are strongly reduced by TSI-13-57, while total MyD88 and actin are unchanged. Bottom: ELISA of serum TNFα, IL-6 and IL-12p40 at 1.5 and 4.5 hours after PAMP injection, with five mice per group. At 4.5 hours TSI-13-57 significantly reduces IL-6 and IL-12p40, whereas TNFα is unaffected.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
We then reconstituted the release mechanism in vitro with three TIR domains. We found that TIRAP restrains MyD88 self-assembly and TRAM relieves that restraint. The purified domains are sufficient to reproduce the behavior seen in cells.
Turbidity assay plotting OD350 over 5 hours for recombinant TIR domains, with lines showing mean and shaded bands showing error. MyD88 TIR alone at 100 µM self-assembles and reaches the highest turbidity. Adding TIRAP TIR at a 1 to 20 ratio (vs MyD88) strongly suppresses assembly. Adding increasing amounts of TRAM TIR, at TIRAP to TRAM ratios of 20:1:1, 20:1:10 and 20:1:20, progressively restores MyD88 assembly towards the level of MyD88 alone.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
We could separate the role of TRAM in the TRIF-arm from its role in the MyD88-arm of TLR signaling based on its localization. Forcing TRAM to the plasma membrane restores myddosome release but abolishes IFN. Endosome-restricted TRAM does the opposite.
Three bar charts showing TNFα, IL-6 and IFNβ secretion measured by ELISA from MyD88-AGF wild-type iBMDMs and ΔTicam2 iBMDMs reconstituted with empty vector, full-length TRAM, or TRAM chimeras carrying the N-terminal plasma membrane targeting sequences of Fyn, Lyn or the TIRAP phosphoinositide-binding motif, stimulated with LTA or LPS. TNFα is restored by all chimeras. IL-6 is restored only by the TIRAP-PBM chimera, which exceeds wild-type levels. IFNβ is restored only by full-length TRAM; all three plasma-membrane-targeted chimeras fail to induce it.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
For releasing myddosomes TRAM acts alongside TIRAP in PI(4,5)P2-rich plasma-membrane microdomains. TRAM mutants, chimeras and CRISPR-Activation showed that the local TIRAP:TRAM ratio, rather than the abundance of either protein, determines whether myddosomes are released.
Fluorescence images of MyD88 localization, MyD88 in grey and nuclei in blue, in MyD88-AGF wild-type iBMDMs and in ΔTicam2 iBMDMs reconstituted with empty vector (EV) or with TRAM WT, G2A, 3X, E180A, C114H or DE88/89AA mutants. The top row is untreated and the bottom row LPS-treated for 1 hour. After stimulation, wild-type cells and cells reconstituted with TRAM WT or E180A show MyD88 assemblies distributed through the cytosol, whereas empty vector, G2A, 3X, C114H and DE88/89AA cells retain MyD88 at the plasma membrane. Scale bars 5 µm.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
Across bacterial infections, TRAM was required for IL-6 but not TNFα production. In infected cells MyD88 again remained at the plasma membrane when TRAM was absent.
Left: three bar charts showing TNFα, IL-6 and IFNβ secretion measured by ELISA from wild-type (white) and Ticam2-/- (orange) BMDMs left uninfected or infected with E. coli (EC) or S. aureus (SA) at MOI 10 for 24 hours. TNFα is comparable between genotypes, IL-6 is strongly reduced in Ticam2-/- cells after both infections, and IFNβ is lost after E. coli but similar after S. aureus infection. Right: fluorescence imaging of MyD88-AGF wild-type and ΔTicam2 iBMDMs infected with E. coli for 3 hours, MyD88 in grey and nuclei in blue. Wild-type cells contain cytosolic myddosomes, whereas MyD88 is retained at the plasma membrane in ΔTicam2 cells, marked by orange arrowheads. Scale bars 5 µm.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
This reflects in transcription: Primary response genes are induced without TRAM, but secondary response genes (e.g. IL-6, IL-12) are not. By RNA-seq, TRAM-deficient macrophages return to baseline by 6 h while WT cells remain activated.
Principal component analysis of RNA-Seq data from wild-type and Ticam2-/- primary BMDMs, untreated or stimulated with LTA or LPS for 2 or 6 hours. PC1 accounts for 81.1% and PC2 for 9.8% of the variance. At 2 hours both genotypes move away from baseline along a shared TLR2/4 early response trajectory. By 6 hours wild-type samples travel further along arrows labelled sustained signaling and TRAM plus TRIF interferon (IFN) response, whereas Ticam2-/- samples curve back towards baseline along an arrow labelled TRAM deficiency.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
The release is what makes TLR signaling last. While WT macrophages sustain NF-κB for hours, TRAM-deficient cells produce a single peak and then terminate signaling early.
Phase plots of p65 nuclear to cytoplasmic ratio, a measure of NF-κB activity, in single cells over 12 hours of stimulation, with 200 cells per condition shown as horizontal traces and activity color-coded from blue (none) to orange (high). Columns show wild-type, ΔMyD88, ΔTirap, ΔTicam1 (TRIF) and ΔTicam2 (TRAM) iBMDMs; the top row is LTA- and the bottom row LPS-stimulation. Wild-type and ΔTicam1 cells sustain NF-κB activity for hours. ΔMyD88 and ΔTirap cells are almost uniformly inactive. ΔTicam2 cells show a bright band of activity only at the start that rapidly returns to baseline.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
Using minute-by-minute co-IPs we found that MyD88 first binds TLR:TIRAP (proto-myddosome), TRAM then releases MyD88 so it can recruit IRAK2/4 and TRAF6 (mature myddosome). Without TRAM, MyD88 stays bound to TLR:TIRAP for hours.
Immunoblots of myddosome immunoprecipitations from MyD88-AGF wild-type and ΔTicam2 iBMDMs stimulated with LPS for 0 to 12 hours, probed for MyD88, TLR4, TIRAP, TRAM, IRAK4, IRAK2, TRAF6 and actin. In wild-type cells, TLR4 and TIRAP associate with MyD88 only within the first minutes, TRAM appears transiently around 30 minutes, and IRAK4, IRAK2 and TRAF6 remain associated for up to 12 hours. In ΔTicam2 cells, TLR4 and TIRAP stay bound for hours, and IRAK and TRAF6 recruitment is delayed, unstable and quickly lost.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
Without TRAM, MyD88 does not move. Proto-myddosomes still nucleate at the plasma membrane but are never released, and no cytosolic myddosomes form. Other adaptor knockouts behaved as expected; ΔTRAM was unique in that myddosome assembly starts but maturation fails.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
To find what builds a myddosome, we performed a genome-wide CRISPR screen, sorting for cells with no myddosomes and no NF-κB activity. We obtained one very unexpected hit: TRAM (Ticam2), the adaptor long exclusively assigned to the MyD88-independent TRIF pathway.
Two rank plots showing gene-level enrichment from MAGeCK robust rank aggregation of the genome-wide CRISPR screen, for LTA- and LPS-stimulated cells sorted for absence of cytosolic myddosomes and NF-κB activation. −log10(MAGeCK score) is plotted against gene rank, with canonical TLR proteins, the oligosaccharyltransferase complex, signaling proteins and controls highlighted in different colors. Rela scores highest in both screens. TRAM is labelled in orange near the top of both rankings, alongside canonical TLR signaling factors MyD88, Traf6, Tirap, Irak4, Tlr2 and Tlr4, whereas Ticam1 (TRIF) ranks far lower.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
We began the study by exploring which cells actually respond to TLR-ligands? Using a side-scatter-based sort we separated macrophages into myddosome-containing responders and myddosome-free bystanders. Only responders switch on the TLR gene program.
Left: Schematic of MyD88-AGF iBMDMs stimulated with PAMPs. Centre left: flow cytometry plot of FSC-W against SSC-A showing the gating strategy, with untreated cells in black and LPS-treated cells (6 h) in blue, separated into SSC-low bystanders and SSC-high responders. Centre right: images of the sorted populations, MyD88 in grey; responders show myddosomes in the cytosol, bystanders do not. Scale bars 10 µm. Right: principal component analysis of RNA-Seq from the sorted populations, in which bystanders separate from responders, and responders cluster with bulk LPS-treated cells.
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Daniel Fisch @danielhfisch.bsky.social · 23/09/2026
Out now @nature.com! We report a new means of cell communication: receptor-free signal transduction. TLRs engage MyD88 for minutes, yet myddosomes signal for hours. TRAM releases myddosomes from TLRs, letting them mature and sustain signaling in the cytosol. rdcu.be/jGdKxErXyAsa Details below:
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Ben Hale @benhale.bsky.social · 21/09/2026
Congrats to Samira Schiefer on her new paper in Science Advances! We analysed 70 natural human IFNB1 variants & found both loss- & gain-of-function IFNβ proteins, revealing unexpected diversity in antiviral immunity. Great effort with Dave Jones & Florence Kwaschik! www.science.org/doi/10.1126/...
science.org
Functional diversity of natural human IFNB1 variants in innate immunity
Human interferon-β (IFN-β), a type I IFN, is critical for effective innate immunity and is also an approved disease-modifying therapeutic. While genetic variations in several components of the type I ...
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Daniel Fisch @danielhfisch.bsky.social · 05/09/2026
Congratulations!!!
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Florian Wollweber @florianwollweber.bsky.social · 05/09/2026
Exciting news! Our @erc.europa.eu StG project ARCHAEAOLOGY got funded 🎉 We’ll combine cryoET/EM, ExM & spatial omics to explore the subcellular organization of Asgard archaea. Job ads coming soon! 🦠 ❄️ 🔬 #TeamTomo #ArchaeaSky #MicroSky
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Payal Yokota @payalyokota.bsky.social · 03/09/2026
Convention would guide us to- one gene, one mRNA, one protein. Venezia and Kane et al suggests challenges that by showing how inflammation pulls distant genes together in 3D space to trans-splice transcripts into hybrids tuning pyroptosis and sepsis outcomes in vivo.👇 www.nature.com/articles/s41...
nature.com
Functional chimeric mRNAs encode proteins in mammalian immunity - Nature
Inflammation induces interchromosomal DNA interactions that bring parent genes into close proximity, facilitating the formation of chimeric mRNAs that encode physiologically relevant, functional prote...
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Carl Zimmer @carlzimmer.com · 27/08/2026
The lab is a mysterious place--where some people have "magic hands," and some experiments just won't work. Here's my story about the unspoken knowledge of science, and the struggle to share it. Gift link: nyti.ms/3UccF7Z
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PLOS Biology @plosbiology.org · 05/08/2026
Graphing and statistically analyzing data is challenging without coding expertise. This Community Page by @avishenoy.bsky.social introduces grafify online, a free, browser app for generating publication-quality graphs and performing statistical tests and comparisons. 🧪 #AcademicSky plos.io/3RpUg6E
Types of graphs and color schemes available in grafify online.
A. Examples of graphs that can be plotted with grafify online. Box and whiskers and violin plots show a line at the median with a box representing the interquartile range (IQR) and whiskers that show 1.5× IQR. Bar graphs and point and error-bar plots show the mean, and the dispersion can be shown as standard deviation (default), standard error of mean, or 95% confidence intervals. Before–after plots show matched data. Numeric XY graphs have numeric variables on both and X- and Y-axes and require a grouping variable that is either categorical (Numeric XY 1) or numeric (Numeric XY 2). B. Categorical (left) and numeric (right) color palettes that are accessible to colorblind individuals
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Daniel Fisch @danielhfisch.bsky.social · 15/05/2026
Congratulation!!! 🎉
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GIMM Institute @gimminstitute.bsky.social · 27/03/2026
Apply until May 3rd, 2026 Join GIMM for a PhD in Biology & Biomedicine (Jan 2027). 🧬 We advance research addressing complex biological questions while providing the foundational and professional skills required for multiple career paths. 🔗 gimm.pt/education-an...
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Daniel Fisch @danielhfisch.bsky.social · 23/12/2025
2/2: TLDR This is a brief video summery of our paper on the cell biology of TLR signaling in innate immune cells. www.nature.com/articles/s41...
nature.com
Molecular definition of the endogenous Toll-like receptor signalling pathways - Nature
Myddosomes, in which MyD88 forms barrel-like scaffold structures for effector protein recruitment and activation, contain proteins that act at all stages and regulate all effector responses ...
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Daniel Fisch @danielhfisch.bsky.social · 23/12/2025
1/2: YouTube knows what I was up to last summer: www.youtube.com/watch?v=UV7r... Many thanks again to @hfspo.bsky.social for inviting me to the Awardees Meeting in Melbourne this July 😁
youtube.com
MOLECULAR DEFINITION OF THE ENDOGENOUS TOLL-LIKE RECEPTOR SIGNALING PATHWAY
YouTube video by HFSP
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MwahahahahahadScientist @mads100tist.bsky.social · 16/11/2025
"Postdoc in the job market"
A headline:

Scientists have discovered a 'third state' between life and death
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bioRxiv Immunology @biorxiv-immuno.bsky.social · 10/11/2025
Truncating RELA variants drive autoinflammation and autoimmunity by impairing the negative feedback control of NF-kB www.biorxiv.org/content/10.1101/202…
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Caetano Reis e Sousa @reisesousalab.bsky.social · 03/11/2025
Delighted that this study is finally out @embojournal.org. A detailed analysis of how DNGR-1/CLEC9A signals to promote cross-presentation of dead cell-associated antigens yet does not activate dendritic cells. www.embopress.org/doi/full/10....
embopress.org
DNGR-1 signalling limits dendritic cell activation for optimal antigen cross-presentation | The EMBO Journal
imageimageDNGR-1 (CLEC9A) is a C-type lectin receptor of type-1 conventional dendritic cells (cDC1s) with an important role in cross-presentation of dead cell antigens. This study explores how it equips cDC1s with the ability to cross-present such ...
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Matteo Carandini @carandinilab.net · 03/11/2025
Europe's MSCA postdoctoral fellowships are now attracting many researchers who would have gone to the US. So the payline has dropped below 10% (~1,600 fellowships to 17k applicants). Super tough situation. One more reason to double the next Horizon budget, as proposed by the EU Commission.
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Pilhofer Lab 🦠❄️🔬 @pilhoferlab.bsky.social · 01/11/2025
🥳 Today marks the start of the lab of former postdoc @florianwollweber.bsky.social at @embl.org Grenoble! www.embl.org/groups/wollw... Florian-- we will miss you dearly and wish you much success! With us, Florian applied multiscale imaging 🔬to Asgard archaea ...👇
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Ivan Zanoni @lozanzi.bsky.social · 31/10/2025
#InflammasomePower! 💥New paper alert💥 How is NLPR3 primed & activated in chronic non-communicable inflammatory diseases? With @oceanedufies.bsky.social &co we show that chronic exposure to oxPAPC drives NRF2 activation that primes & activates NLRP3 sustaining #atherosclerosis in mice & humans! 👇👇👇
biorxiv.org
Chronic sensing of host-derived lipids is an all-in-one signal that primes and activates NLRP3.
Activation of the NLRP3 inflammasome leads to the production of bioactive interleukin (IL)-1β fostering atherosclerosis. The current dogma is that NLRP3 must be first primed by microbial stimuli, know...
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Nils Burger @nilsburger.bsky.social · 16/10/2025
Excited to share that we are hiring a research assistant for my lab at UTSW in Dallas (www.nils-burger-lab.com). Please share with your mentees. utsw.taleo.net/careersectio... We are also looking for motivated, curious, and enthusiastic postdoctoral fellows!
utsw.taleo.net
Research Assistant I, Pharmacology
Click the link provided to see the complete job description.
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RedPen/BlackPen @redpenblackpen.bsky.social · 15/10/2025
What's in yer basement?
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Avinash Shenoy @avishenoy.bsky.social · 28/09/2025
📢A new effector-triggered immunity role for GBP1! No place for pathogens to hide now that GBP1 can detect extracellular pathogens that hijack the actin cytoskeleton. Surprise*suprise* GBP1 does this independently of LPS! 🧵 biorxiv.org/content/10.1...
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Jonathan Kagan @jkagan1.bsky.social · 26/09/2025
meta-PAMPs: A new class of immunostimulatory molecules that control the ability of bacterial factors to operate as PAMPs. Jordan Jastrab reveals that wall teichoic acid (WTA) is a key regulator of inflammasomes and interferons during infection. www.biorxiv.org/content/10.1...
biorxiv.org
Wall teichoic acid is required for DNA-triggered innate immune receptor activation by Staphylococcus aureus
Receptors that stimulate inflammation are commonly activated by ligands that are buried within microbial cells. The mechanisms that facilitate immunostimulatory ligand release from microbes during inf...
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Jordan Jastrab @jordanjastrab.bsky.social · 26/09/2025
Thrilled to share my work from the @jkagan1.bsky.social lab! TLDR: O-acetylation and wall teichoic acid compete for space on S. aureus peptidoglycan. This competition regulates DNA release, and thus immune receptor activation, during infection! www.biorxiv.org/content/10.1... #microsky #immunosky
biorxiv.org
Wall teichoic acid is required for DNA-triggered innate immune receptor activation by Staphylococcus aureus
Receptors that stimulate inflammation are commonly activated by ligands that are buried within microbial cells. The mechanisms that facilitate immunostimulatory ligand release from microbes during inf...
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Thiago Carvalho @cyrilpedia.bsky.social · 07/09/2025
Tools: "Here, we developed a class of highly sensitive genetically encoded STAT biosensors, termed STATeLights, which allowed direct and continuous detection of STAT activity in live cells with high spatiotemporal resolution." www.nature.com/articles/s41...
nature.com
Real-time visualization of STAT activation in live cells using genetically encoded biosensors - Nature Chemical Biology
The signal transducer and activator of transcription (STAT) family proteins are attractive drug targets but tools to monitor their activation are lacking. Now, STAT biosensors have been developed for ...
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Dr Rebecca Coll @colllab.bsky.social · 02/09/2025
So excited that this study is now online! It's been a pleasure to work with BioAge Labs and our collaborators on this story. We characterise a new class of #NLRP3 inhibitors with translational potential for so many inflammatory diseases.
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Simon Kuestenmacher @simongerman600.bsky.social · 20/08/2025
Guilty as charged.
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Ivan Zanoni @lozanzi.bsky.social · 21/08/2025
#InterferonPower! Thrilled for our latest work @cp-cell.bsky.social! With @danielboehmer.bsky.social, we dug into tons of papers & created what we hope will be a go-to resource for immunologists & non-immunologist about type I, II, III (& IV😉) #interferons! Free👉 authors.elsevier.com/a/1leKGL7PXu...
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Adam Braithwaite @braithwaiteat.bsky.social · 03/07/2025
Stimulating immune cells in vitro is a common experimental lab model. We profiled 150K blood immune cells treated with 11 different stimuli to compare the effects. The data are freely available for researchers. See the preprint for our findings and to access the data 🧪 doi.org/10.1101/2025...
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Waggoner Lab @labwaggoner.bsky.social · 12/08/2025
Divergent TIR signaling domains in TLR7 and TLR9 control opposing effects on systemic autoimmunity @jclinical-invest.bsky.social www.jci.org/articles/vie...
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Jonathan Kagan @jkagan1.bsky.social · 30/07/2025
Everything you wanted to know about oxidized lipids & immunity, in one review. We explain how oxidized lipids impact inflammation, dendritic cells, anti-tumor immunity and more. Kudos to Joon Choi for his exceptional work. royalsocietypublishing.org/doi/10.1098/...
royalsocietypublishing.org
Oxidized phospholipid damage signals as modulators of immunity | Open Biology
Damage-associated molecular patterns (DAMPs) are self-derived molecules released during tissue damage that influence immune responses. Phospholipids, essential to cell membranes and lung surfactants, ...
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Chanda Prescod-Weinstein 🌌 @chanda.blacksky.app · 27/07/2025
Zotero has changed my life for the better. What a marvelous piece of software.
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