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. 2020 Mar 11;12(3):655. doi: 10.3390/cancers12030655

Figure 6.

Figure 6

siBNIP3L inhibited cancer stemness induced by HBx-expressing in MHCC-97H cells. MHCC-97H cells without or with HBx-expressing were transiently transfected with pcDNA3.1 or pcDNA3.1-HBx (1 μg/mL) and siBNIP3L or siNC (50 nmol/L). (A) Representative images of the immunofluorescence co-staining for MitoTracker (red), BNIP3L (blue), and LC3B (green). The profiles of representative lines trace the intensities of fluorescence signals. Fluorescence curves with line intensity profile generated by Zen 2012 software were shown. Scale bar represents 10 μm. (B) The BNIP3L-dependent mitophagy-related proteins in cytoplasmic (Cyto) and mitochondrial (Mito) fractions. (C) The expression levels of cancer stemness-related proteins. The gray value of band was assessed by image-pro plus 6.0. The relative expression level was shown. (D) The mRNA levels of cancer stemness-related genes. The target gene transcription was normalized to ACTB. (E) The self-renewal capacity was measured by colony formation assay. (F) Percentage of the sorted SP cells (R1 gate) were detected by FCM (Left). Quantitative results were shown in bar graph (Right). SP: side population. R1 gate represented SP cells. * P < 0.05 as compared with pc3.1 group. # P < 0.05 as compared with pc3.1-HBx group. pc3.1: pcDNA3.1 transfection without HBx-expressing. pc3.1-HBx: pcDNA3.1-HBx transfection with HBx-expressing.