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. 2016 Apr 15;11(6):3702–3708. doi: 10.3892/ol.2016.4442

Figure 2.

Figure 2.

Activity and expression of YAP is regulated by c-Jun/c-Fos. (A and B) Overexpression of c-Jun/c-Fos upregulated YAP protein expression as shown by (A) representative western blotting images and (B) quantification of YAP, c-Jun and c-Fos in human HCC Bel-7402 cells with or without c-Jun/c-Fos overexpression. (C) c-Jun/c-Fos upregulated YAP mRNA levels. (D and E) Knockdown of c-Jun/c-Fos reduced YAP protein expression as shown by (D) representative western blotting images and (E) quantification of YAP, c-Fos and c-Jun in Bel-7402 cells with or without c-Jun/c-Fos-knockdown. (F) YAP activity was regulated by c-Jun/c-Fos. The pUAS-Luc-TEAD-Gal4 luciferase reporter systems were transfected into human HCC Bel-7402, SMMC-7721 or Huh7 cells with or without c-Jun/c-Fos-knockdown or overexpression (transfected with 0.5–1.5 µg plasmids). (G) Endogenous cellular YAP expression was increased by overexpression of c-Jun/c-Fos, as measured by immunofluorescence in Bel-7402 cells transfected with exogenous c-Fos-FLAG or c-Jun-FLAG-expressing plasmids. Arrows indicate cells with c-Jun/c-Fos transfection, while asterisks indicate cells without successful transfection. Scale bar, 20 µm. Data are presented as the mean ± standard deviation of three independent experiments. Empty and GFP-sh groups were arbitrarily set to 100%. *P<0.05. YAP, yes-associated protein; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; sh, small hairpin; GFP, green fluorescent protein; TEAD, TEA domain; Luc, luciferase; DAPI, 4′,6-diamidino-2-phenylindole; HCC, hepatocellular carcinoma.