Extended Data Fig. 1.

GCDH inhibition promotes apoptosis of melanoma cells (A) UACC903 and 1205LU melanoma cells were transduced with siRNAs targeting AASS, AADAT, DHTKD1, GCDH, or ECHS1 or with a control sequence using Jetprime for 96 hr, followed by an analysis of cell viability. (B) Various melanoma cells were transfected with siRNAs targeting AASS, AADAT, DHTKD1, GCDH, or ECHS1 or with a control sequence for 72 hr, and then lysates were subjected to western analysis with indicated antibodies. (C) Cell growth after GCDH KD with two independent siRNAs for 0–96 hr in UACC903 or 1205LU cells. Proliferation was analyzed by counting cells at indicated time points. (D) UACC903 or 1205LU cells were transfected with GCDH siRNA for 72 hr, and then lysates were subjected to western analysis with indicated antibodies. (E) Viability assay of control or GCDH KD A375 cells treated with the caspase inhibitor Z-VAD(OH)-FMK (10μM for 48 hr). (F) A375 cells transfected with GCDH siRNA were grown in the indicated growth medium, and then viability was determined 96 hr-post transfections. (G) Measurement of basal, maximum oxygen consumption rate (OCR), and spare respiratory capacity using Agilent Seahorse XF Analyzers. A375 cells were transfected with GCDH siRNA for 48 hours before analysis. (H) Viability assay of immortalized H3A cells, 96 hr after transfection with indicated constructs. Viability was measured by quantifying crystal violet staining. (I) Immortalized H3A cells were transfected with siRNAs targeting AASS, AADAT, DHTKD1, GCDH, or ECHS1 or with a control sequence for 72 hours, then lysates were subjected to western analysis with indicated antibodies. Data are representative of three experiments for (B, D, and I) and presented as the mean values ± SD of n = 3 (A, C, and H), n= 4 (E and F) independent experiments and mean values ± SD of n = 6 technical replicates for (G). Each p value is adjusted to account for multiple comparisons. Statistical significance (indicated p-value or ns- not significant w.r.t control) was calculated using One-way ANOVA for (B, E ,F and H) and two-way ANOVA for (C). Statistics source data for Extended data figure 1 can be found in Source data files - Extended data figure 1 and uncropped scans of all blots and gels for Extended data figure 1 can be found in Source data files for blots - Extended figure 1.