Fig. 3. METTL16 promotes cuproptosis by targeting FDX1.

a–e MeRIP-seq analysis of m⁶A regulation between METTL16 and FDX1 in HGC-27 cells. a Workflow for methylated RNA immunoprecipitation sequencing (MeRIP-seq). b Specific m⁶A motif analysis in stable Ctrl-Sh and METTL16-Sh cell lines. c Significant 15 GO terms of METTL16-related genes identified between Ctrl-Sh mRNAs and METTL16-Sh mRNAs containing m⁶A modifications. The green arrow indicated the specific terms that were related to cuproptosis. Enrichment Scores were calculated using chi-square test. d Venn plot showing the intersection of downregulated mRNAs and downregulated m⁶A in METTL16-Sh cells. The gene list showed the cuproptosis-related genes in the intersection. e qRT-PCR analysis of FDX1 in Ctrl and shMETTL16-Sh AGS and HGC-27 cells (n = 3). HGC-27: P = 0.0023. AGS: P = 0.0064. f, g Western blot analysis of METTL16 and FDX1 expressions in Ctrl and METTL16-Sh cells (f) or METTL16-knockout (KO-1, KO-4, KO-6) (g) HGC-27 cells. h Western blot analysis of METTL16 and lipoylated DLAT expression in METTL16-knockdown and control HGC-27 cells. i–k Growth survival was analyzed by CCK8 assay. i, j FDX1-overexpression reversed the recovery of growth inhibition induced by METTL16-knockdown (i) or knockout (j) cell lines (n = 3). P = 0.00668 (Sh1) and 0.00984 (KO-1). k FDX1-knockdown rescued the growth inhibition induced by METTL16-overexpression cells. Stable control or METTL16-overexpression cell lines transfected with or without FDX1 silencing plasmids were treated with indicated different concentrations of elesclomol-Cu (ratio = 1:1) for 72 h (n = 3). P = 0.00617. Statistical data presented in this figure show mean values ± SD of three times of independent experiments. Statistical significance was determined by Two-tailed t test, *P < 0.05, **P < 0.01. Source data are provided as a Source data file.