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methylcobalamin  (MedChemExpress)
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MedChemExpress methylcobalamin
a Procedure of terbium (Tb 3+ )/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 <t>(methylcobalamin,</t> 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 K d 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.
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a Procedure of terbium (Tb 3+ )/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 K d 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.

Journal: Nature Communications

Article Title: Methylcobalamin protects against liver failure via engaging gasdermin E

doi: 10.1038/s41467-024-54826-6

Figure Lengend Snippet: a Procedure of terbium (Tb 3+ )/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 K d 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.

Article Snippet: Methylcobalamin (HY-B0586), hydroxocobalamin (HY-B2209A), adenosylcobalamin (HY-112790), cyanocobalamin (HY-B0315), L-methionine (HY-N0326), tetrahydrofolic acid (HY-14520), dimethyl fumarate (HY-17363), Z-DEVD-FMK (HY-12466), cisplatin (HY-17394), acetaminophen (HY-66005), N-acetylcysteine (HY-B0215) were purchased from MCE (NJ, USA).

Techniques: Fluorescence, Inhibition, Positive Control, Lactate Dehydrogenase Assay, Recombinant, Activity Assay, Binding Assay