cflip  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: A TAK1 Cytokine Toxicity Checkpoint Controls Anti-Cancer Immunity

doi: 10.1101/2025.05.09.652721

Figure Lengend Snippet: (A) Analysis from Lawson et al , highlighting CFLAR as an immune evasion gene across multiple cell lines. (B) Time-course immunoblot showing cFLIP (FLIP L isoform) expression in wild-type and TAK1-deficient B16F10 cells treated with IFNγ and TNF for the indicated durations (0-8 hours). (C) Western blot of FLIP L expression after 4-hours stimulation with IFNγ, TNF, or their combination in WT and TAK1-deficient B16F10 cells. (D) cFLIP protein levels in WT and TAK1-deficient B16F10 cells treated with TNF (4 hours) with or without proteasome inhibitor MG132 (5 µM). (E-F) Similar cytokine-induced downregulation of cFLIP (FLIP L and FLIP s ) observed in HeLa cells (E) treated with IFNy (100 ng/mL), TNF (2 ng/mL) or/and TAKi (5 µM), and where indicated (F), in cells treated with Emricasan (5 µM). All these indicate a conserved effect of TAK1 inhibition across cell types. (G) Flow cytometry quantification of PI⁺ (dead) cells following 24-hours treatment with IFNγ and TNF in control, TAK1-deficient, cFLIP-deficient, or double-deficient B16F10 cells.

Article Snippet: Membranes were then probed overnight at 4°C with the following primary antibodies: CASP3 (CST, Cat#9662), cleaved CASP3 (CST, Cat#9661T), CASP8 (CST, Cat#4927T), cleaved CASP8 (CST, Cat#9429T), CASP8, (AdipoGen, Cat#AG-20B-0057-C100), RIPK1 (CST, Cat#3493S), PARP (CST, Cat#9542T), STAT1 (CST, Cat#9172T), TAK1 (CST, Cat#5206S), CFLIP (CST, Cat#56343), A20 (Santa-Cruz, Cat#sc-166692), FADD (CST, Cat#2782), TRADD (Santa-Cruz, Cat#sc-46653), ACTIN (Proteintech, Cat#66009-1-Ig) and GAPDH (Invitrogen, Cat#437000).

Techniques: Western Blot, Expressing, Inhibition, Flow Cytometry, Control

Journal: Cell Death Discovery

Article Title: Regulation of ER stress-induced apoptotic and inflammatory responses via YAP/TAZ-mediated control of the TRAIL-R2/DR5 signaling pathway

doi: 10.1038/s41420-025-02335-w

Figure Lengend Snippet: A A549 cells were plated on plastic or collagen-coated polyacrylamide gels with different rigidity as described in Material and Methods and treated with thapsigargin 200 nM during the indicated time points. Protein expression was analyzed by western blotting. B A549 cells were transfected with a scrambled oligonucleotide (SC) or siRNA YAP/TAZ (Y/T). Thirty hours post-transfection, cells were treated with thapsigargin and protein expression was analyzed by western blotting. C A549 cells were transfected with two different siRNA against both YAP and TAZ. 48 h post-transfection, cFLIP mRNA levels were assessed by qPCR (left panel). In right panel, A549 cells were transfected with siRNA targeting YAP/TAZ, and 30 h after transfection cells were treated overnight with thapsigargin, and cFLIP mRNA levels were assessed by qPCR. Data show the mean ± SD of from three independent experiments ( D ) A549 cells were transfected with siRNA oligonucleotides targeting both cFLIP isoforms. After 6 h cells were treated with thapsigargin for 24 h. Cell death was analyzed by subG1 quantification. Data show the mean ± SD of at least three independent experiments. E A549 pBabe or A549pBabe-FLIPL were transfected with siRNA SC or siRNA Y/T. After 30 h, cells were treated with thapsigargin 200 nM during 24 h. Caspase-8 (C8) activation was analyzed by western blotting. F Cells were treated as in ( E ) and cell death was analyzed by quantification of subG1 population. Data represent mean ± SD from three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Unpaired Ttest ( C ), Two-way ANOVA with Tukey´s multicomparisons test ( D , F ).

Article Snippet: siRNAs against YAP, TAZ, cFLIP, and non-targeting scrambled oligonucleotide (Supplementary Information section) were synthesized by Merck/Sigma Aldrich.

Techniques: Expressing, Western Blot, Transfection, Activation Assay

Journal: Cell reports

Article Title: Non-canonical autophosphorylation of RIPK1 drives timely pyroptosis to control Yersinia infection

doi: 10.1016/j.celrep.2024.114641

Figure Lengend Snippet:

Article Snippet: cFLIP Cell Signaling Technology , , Cat#: 56343; RRID: AB_2799508.

Techniques: Virus, Recombinant, Staining, Electroporation, CRISPR, Enzyme-linked Immunosorbent Assay, In Situ, Gene Expression, Sequencing, Mutagenesis, Software