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. 2016 May 9;7(26):39694–39704. doi: 10.18632/oncotarget.9233

Figure 3. The effects of Pinin on clonogenicity and proliferation in HCC cells.

Figure 3

a, c. shRNA-mediated silencing of Pinin expression in human HCC cell lines. Western blot and q-RT-PCR detected the expression of Pinin in SNU449 and BEL7402 cells. b, d. Cells with or without knockdown of Pinin were cultured for the days as indicated and cell growth was evaluated by trypan blue staining (n=3, mean ± SD, t-test, *P<0.05, **P<0.01 vs. shRNA CTR). e, f. Colony formation assay was used to measure the clonogenicity of SNU449 and BEL7402 cells with or without knockdown of Pinin. Cells were grown in the same condition. All dishes were fixed, stained and photographed at the same time. The number of untreated cells were set as 100% (n=3, mean ± SD, t-test, **P<0.01 vs. shRNA CTR). g, h. SNU449 and BEL7402 cells with or without knockdown of Pinin were stained with EdU. The nuclei were also stained by Hoechst 33342. The percentage of cell proliferation was expressed as the ratio of EdU positive cells to total Hoechst 33342 positive cells. The number of untreated cells were set as 100% (n=3, mean ± SD, t-test, **P<0.01 vs. shRNA CTR). i-k. Colony formation assay was used to measure the clonogenicity of SNU449 and BEL7402 cells with or without overexpression of Pinin. Cells were grown in the same condition. All dishes were fixed, stained and photographed at the same time. Western blot detected the protein level of Pinin using the antibodies as indicated. The number of untreated cells was set as 100%. (n=3, mean ± SD, t-test, *P<0.05 vs. pCDH-flag).