Fig. 3. Mechanism investigation of pRNCThioether+DEA mediated B16F10 ICD.
a, b Representative images of pRNCThioether+DEA mediated co-localization with mitochondria and ER. MitoTracker green (a) and ER Tracker green (b) were stained with mitochondria and ER, respectively. Red color represented Cy5-pRNCThioether+DEA, and blue color represented DAPI which were stained by Hoechst 33342. PCC = Pearson’s correlation coefficient. Scale bar = 10 μm. c Volcano map of expressed differentially genes in pRNCThioether+DEA treated cells. d Heatmap depicting relative transcript levels of expressed differentially genes in pRNCThioether+DEA or PBS treated cells (n = 3 independent experiments). e KEGG pathway enrichment analysis of expressed differentially genes for cells after pRNCThioether+DEA treatment. f, g Enrichment of organismal systems pathways (f) and metabolism pathways (g) in pRNCThioether+DEA or PBS treated cells (n = 3 independent experiments). h Intracellular ROS level after different treatments via flow cytometer characterization (n = 3 independent experiments). i Intracellular mtROS level after treatments via CLSM characterization. Scale bar = 20 μm. j Western blot of p-PERK, p-elf2α, ATF4, cleaved caspase-1, N-GSDMD, MLKL expression after treatments. β-actin was used as internal control. Experiments in (a), (b), (i), and (j) were repeated three times independently with similar results.