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. 2022 May 23;10:840171. doi: 10.3389/fcell.2022.840171

FIGURE 8.

FIGURE 8

TMAO activates ILK/AKT/mTOR signaling via Postn in Huh7 cells. (A) The translation levels of POSTN, ILK, AKT, p-AKT, mTOR and p-mTOR measured by western blotting in Huh7 cells treated with 50 ng/ml TNF-α combined with 50 μM TMAO or 50 ng/ml TNF-α alone. Grayscale values were analyzed; data are mean ± SD (n = 3 per group); Student’s t-test; ***, p < 0.001. (B) POSTN translation levels in Huh7 cells infected with pseudovirus expressing sh-POSTN (sh1, sh2, sh3) plasmids were compared with the Huh7 cells infected with pseudovirus expressing scramble plasmids measured by western blotting. Grayscale values were analyzed; data are mean ± SD (n = 3 per group); one-way ANOVA with Bonferroni’s test; ***, p < 0.001. (C) CCK-8 assay of the effect of knocking down POSTN on the proliferation of Huh7 cells; data are mean ± SD (n = 3 per group); one-way ANOVA with Bonferroni’s test; ***, p < 0.001. (D) Transwell assays of the effect of knocking down POSTN on the invasive abilities of Huh7 cells. Scale bars = 100 μm; Data were analyzed by ImageJ; data are mean ± SD (n = 3 per group); one-way ANOVA with Bonferroni’s test; ***, p < 0.001. (E) Western blots of the effects of knocking down POSTN on the protein levels of Postn, ILK, AKT, p-AKT, mTOR and p-mTOR in Huh7 cells. Grayscale values were analyzed; data are mean ± SD (n = 3 per group); one-way ANOVA with Bonferroni’s test; ***, p < 0.001.