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

Figure 2.

Figure 2

HBx promoted cancer stemness phenotype of the HCC cells. (A–E) The two HCC cell lines without or with HBx-expressing were transiently transfected with pcDNA3.1 or pcDNA3.1-HBx (0.5, 1 μg/mL) for 8 h and then restored culture for another 48 h. (A,B) The expression levels of cancer stemness-related proteins (A), and the mRNA levels of cancer stemness-related genes (B) in two HCC cells without or with HBx-expressing. The target gene transcription was normalized to ACTB. (C) Percentage of the sorted SP (R1 gate) in two HCC cells without or with HBx-expressing (Left). Quantitative results were shown in bar graph (Right). SP: side population. R1 gate represented SP cells. (D,E) The self-renewal capacity was analyzed by anchorage-independent growth assay (D) and colony formation assay (E) in two HCC cells without or with HBx-expressing. Magnification (200×). (F) The expression levels of cancer stemness-related proteins in two HCC cells transiently transfected with pGEM, pGEM-HBV, and pGEM-HBV X null plasmids. The gray value of band was assessed by image-pro plus 6.0. The relative expression level was shown. * P < 0.05 as compared with Huh7-pc3.1 group. # P < 0.05 as compared with MHCC-97H-pc3.1 group. pc3.1: pcDNA3.1 transfection without HBx-expressing. pc3.1-HBx: pcDNA3.1-HBx transfection with HBx-expressing.