Fig. 4

Pt@PCN-Cu enhances PD-L1 expression through an HK2-dependent mechanism. (A) PANC-1 cells were treated with Pt@PCN-Cu for 24 h and PD-L1 expression was assessed. (B) PANC-1 cells were treated with Pt@PCN-Cu for 24 h in the presence or absence of CHX. (C) PANC-1 cells were cultured with Pt@PCN-Cu for 24 h in the presence or absence of actinomycin D. (D) The qPCR analysis of CD274 mRNA expression. (E) Volcano plot showing differential gene expression between PBS group and Pt@PCN-Cu group. (F) Enrichment pathway analysis identifying the most significantly altered signaling pathways. (G) Enrichment pathway analysis highlighting key pathways. (H) Heatmap showing differentially expressed genes. (I) Western blot analysis of PD-L1 expression in PANC-1 cells stably expressing control shRNA or HK2 shRNA, treated with or without G-6-P for 12 h. (J) Western blot analysis of PD-L1 expression in PANC-1 cells stably expressing control shRNA or HK2 shRNA and cultured in the presence of Pt@PCN-Cu. (K-M) Mitochondrial and cytosolic fractions were isolated and western blot analysis was performed to evaluate HK2 and VDAC1 expression in each fraction. (N, O) Co-immunoprecipitation assay demonstrating the interaction between HK2 and VDAC1. Data are presented as mean ± SD. Statistical significance was determined using t-test, with **P < 0.01