Extended Data Fig. 3: Inhibition of palmitoylation suppresses GSDMD processing.
From: A palmitoylation–depalmitoylation relay spatiotemporally controls GSDMD activation in pyroptosis
a, Primary BMDMs were primed with LPS (100 ng/ml) for 3 hours followed by treatment with 2-BP (50 μM) for 0.5 hours before stimulation with Nig (10 μM) for 2 hours. Western blot analyses were performed with anti-GSDMD antibodies. b, BMDMs were primed with LPS (100 ng/ml) for 3 hours followed by treatment with 2-BP (50 μM) for 0.5 hours before stimulation with ATP (2 mM) for the indicated times. Western blot analyses were performed with anti-GSDMD antibodies. c, BMDMs were pre-treated with 5z7 (200 nM) with or without 2-BP (50 μM) and Nec-1s (10 μM) for 0.5 hours followed by treatment with LPS (40 ng/ml) for the indicated times. Western blot analyses were performed with anti-GSDMD antibodies. d, Primary BMDMs were primed with LPS (100 ng/ml) for 3 hours followed by treatment with 2-BP (50 μM) for 0.5 hours before stimulation with Nig (10 μM) for the indicated times. Western blot analyses were performed with anti-NLRP3 antibodies. e, Primary BMDMs were treated with LFn-Rod (100 ng/ml) plus protective antigen (1 µg/ml) with or without 2-BP (50 μM) for 1.5 hours. Volcano plots of RNA-seq showing fold-change and P-value for the comparison of BMDMs treated with 2-BP versus Veh (n = 3) with cutoffs set at fold change (FC) > 2 or < 0.5, and P-value < 0.05 were shown. Representative genes associated with pyroptosis are highlighted. Unpaired two-tailed Student’s t-test.
