Fig. 6: Analysis of mitophagy regulation and cell demise mechanisms by RNA sequencing after treatment with SHC4H+hv under hypoxia (1% O₂).

a Bubble diagram of the DEGs enriched in the GO pathway (False Discovery Rate, FDR < 0.05), n = 3 independent experiments. b GSEA of the DEGs in B16 cells after treatment with SHC4H followed by irradiation (p-val<0.05), n = 3 independent experiments. A: Positive regulation of the execution phase of apoptosis; B: Regulation of cyt (c) release from mitochondria. C: Regulation of mitochondrial membrane potential. D: Cytochrome c oxidase activity. E: Positive regulation of intrinsic apoptotic signaling pathway. NES: normalized enrichment score. c GSEA of the DEGs in MCF-7 cells after treatment with SHC4H followed by irradiation (p-val<0.05), n = 3 independent experiments. A: DNA adduct; B: Glutathione metabolism; C: Reactive oxygen species; D: Mitophagy; E: Necroptosis. d Heatmap of genes associated with the mitophagy and cell death mode (Apoptosis or Necroptosis) in B16 and MCF-7 after different treatments. The red color represents upregulation, and the blue color indicates downregulation, n = 3 independent experiments. e PPI (protein-protein interaction) network of identified differentially expressed genes after treating SHC4H+hv in B16 and MCF-7 cells (n = 3 independent experiments). f The amount of intracellular cytochrome-c after different treatments in B16 and MCF-7 cells (n = 5 independent experiments; mean values  ±  SD). g The expression of key proteins for apoptosis in B16 cells, including cleaved-Caspase-3 (cCasp3), Bcl-2, after different treatments. Four independent experiments were performed and representative results are shown. h After different treatments, the expression of key proteins for necroptosis in MCF-7 cells, including p-Mlkl and p-Ripk3. Three independent experiments were performed and representative results are shown. i The levels of intracellular TNF after different treatments in MCF-7 cells (n = 4 independent experiments; mean values  ±  SD). Statistical differences were determined by One-way ANOVA with a Tukey post-hoc test. Source data are provided as a Source Data file.