Figure 3. IRF-1 has anti-proliferative and pro-apoptotic effect on HCC cells.

A) IRF-1 and CXCR3 mRNA expression are determined by qRT-PCR in tumor from HCC patients (n=10). B) Correlative analysis of IRF-1 and CXCR3 mRNA expression in HCC patients is based on TCGA database (n=360). Each data point (A & B) represents one patient. CXCR3 mRNA (C) and protein (D) expression are respectively measured by qRT-PCR and western blot in Hepa1–6 cells and Huh-7 cells infected by mouse / human AdIRF-1 (50 MOI) for 48 h. E) IRF-1 mRNA (upper) and total protein (lower) expression are respectively analyzed by qRT-PCR and western blot in HCC cells stably overexpressing IRF-1 (IRF-1) vs. negative control (NC) and wild type (WT). The qRT-PCR data are presented as mean ± SEM and shown statistical difference from three independent experiments. F, G) CXCL10 and CXCR3 mRNA (F) and protein (G) expression are respectively determined by qRT-PCR and immunofluorescent staining in IRF-1 vs. NC (×400 magnification). H, I) The statistical summary of apoptotic Hepa1–6 cells rate (H) and representative images (I) of FACS assay for apoptotic (Annexin V+PI+) cells rate in Hepa1–6 cells expressing IRF-1 vs. NC. Each data point (C, F, & H) represents an independent experiment. Data represent mean ± SD. J) MTT assay for cells expressing IRF-1 vs. NC at different time points. The data are presented as the means ± SEM of six samples and are representative of two independent experiments with similar results. K) Western blot analysis for cleaved PARP in Huh-7 cells infected by human AdIRF-1 (50 MOI) for 48 h. Western blot and immunofluorescent results shown are representative image from two independent experiments. *p<0.05, **p<0.01.