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

FIGURE 5.

FIGURE 5

Screening of the functional gene in the combination strategy against HCC. (A) Heatmap of the RNA sequencing results. Changes in the relative abundance of proteins and components from Huh7 tumor-bearing mice treated with RT with or without AT-MSCs (n = 2–3 per group). (B–C) Venn diagram and heatmap showing 76 genes differentially expressed between the CTRL and RTM, RT and RTM, and MSC and RTM groups. (D) Functional interaction sub-network constructed using module genes. (E) Western blot analysis demonstrating IFITM1 expression in HCC xenograft tumor samples. β-Actin was used as a loading control. (F) Detection of IFITM1 mRNA expression in xenograft tumor samples by qRT-PCR after RT with or without AT-MSCs treatment. Fold change expression was calculated with normalization to GAPDH as the internal control gene. Each value represents the mean ± SD from three independent experiments (∗P < 0.05, ∗∗∗P < 0.001). (G) Representative images of IHC staining of the proliferative marker Ki-67 (top) and IFITM1 and TUNEL staining (bottom) in xenograft tumors (n = 3–5 tumors per group, five images per tumor. Scale bar = 100 μm). (H) The percentages of Ki- 67-, IFITM1-, and TUNEL-positive cells are summarized in the bar chart (∗∗P < 0.01, ∗∗∗P < 0.001). (I) Protein expression of IFITM1 in HCC cells and the immortalized liver cell lines LO2 was detected by Western blot analysis. β-actin was used as a loading control. (J) Detection of IFITM1 mRNA expression in HCC cell lines and LO2 by qRT-PCR. The results are expressed as the means ± SD of three independent experiments (∗∗P < 0.01). (K) Detection of secreted IFITM1 level in HCC cell lines and immortalized liver cell lines. (L) Representative image of immunofluorescence staining of IFITM1 (green) in indicated cells; nuclei are stained with DAPI (blue).