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. 2019 Nov 12;7:267. doi: 10.3389/fcell.2019.00267

FIGURE 6.

FIGURE 6

IFITM1 is identified as a potential HCC-oncogene and involves in the synergy treatment. (A,B) Detection of IFITM1 protein and mRNA expression in Huh7 cells treated with RT with or without AT-MSCs by Western blot analysis and qRT-PCR. β-Actin was used as a loading control. (C) Representative image of immunofluorescence staining of STAT3 (red) and IFITM1 (green) in the indicated cells; nuclei are stained with DAPI (blue). (D,E) HCC cell lines were transduced with shRNAs targeting IFITM1, and the expression level of IFITM1 was assessed by Western blot analysis and qRT-PCR using β-Actin and GAPDH as loading controls, respectively. The results are expressed as means ± SD of three independent experiments. (F) The viability of Huh7 and HepG2 cells transfected with shRNAs were assessed by CCK8 assay for 24 h. Data represent the means ± SD of three independent experiments (∗∗P < 0.01, ∗∗∗P < 0.001). (G,H) Huh7 cells transfected with shRNAs targeting IFITM1 (shIF-1 or shIF-2) or negative control (NC) shRNA were cultured on ultra-low attachment plates to determine the sphere-formation ability. Representative images are shown. The results are expressed as the means ± SD of three independent experiments (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001). (I,J) A colony-formation assay showed decreased tumorigenicity of Huh7 cells transfected with shIF-1 or shIF-2 compared with NC shRNA in vitro. The graph shows the mean integrated density of colonies normalized by 100% (∗∗∗P < 0.001).