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. 2019 Aug 19;20(16):4048. doi: 10.3390/ijms20164048

Figure 6.

Figure 6

SIRT1 overexpression leads to downregulation of sorafenib-induced p53 signaling. HUH7 cells were stimulated with sorafenib (5 µM). SIRT1 was overexpressed in HUH7 cells by electroporation. (A) Left: Densitometric analysis of SIRT1 was done using ImageJ (NIH) [17]. SIRT1 was normalized to mean values of GAPDH, which served as a loading control. Right: Acetylated Lys382-p53, total p53, poly (adenosine diphosphate ribose) polymerase 1 (PARP-1), and p53 upregulated modulator of apoptosis (PUMA) was detected by Western blot analysis, GAPDH served as a loading control. One representative blot out of three experiments is shown. Densitometric analysis of (B) acetylated Lys382-p53, (C) total p53, and (D) PUMA was done using ImageJ (NIH). Solvent control (DMSO) was set =1, indicated by a line. One representative blot out of three experiments is shown. Data are shown as mean ± SEM. Sirt1-FLAG: overexpression plasmid for Flag-tagged SIRT1.