Fig. 1: Methylcobalamin is a specific and direct inhibitor of GSDME.
From: Methylcobalamin protects against liver failure via engaging gasdermin E
a Procedure of terbium (Tb3+)/dipicolinic acid (DPA) fluorescence liposome leakage assay. b Percentage of inhibition on 829 compounds, assayed at 100 μM. Cutoff was 70% inhibition. Dimethyl fumarate (DMF) was used as a positive control. Each point is an individual inhibitor (n = 1). c LDH release assay for cytotoxicity of HepG2 cells pretreated with 9 screening hits at 100 μM before challenged with TNFα/cycloheximide (CHX) for 16 h to induce GSDME-mediated pyroptosis. d Chemical structure of compound 19A11 (methylcobalamin, MeCbl). e 25 ~ 100 μM MeCbl or DMF were treated with 0.2 μM recombinant active-caspase-3 for activity assay. Z-DEVD-FMK was used as positive control. f MST measurements of the binding of 0.2 μM recombinant caspase-3, GSDME, GSDMD with MeCbl or positive control. Values of Kd are listed in the graphs. Data are mean ± SEM (n = 3) of at least two independent experiments for bar charts. Analysis was done using one-way ANOVA (c, e). p < 0.05 is considered significant and p values are provided in graphs, n.s = nonsignificant. All groups were compared to indicated vehicle. Source data are provided as Source Data file.
