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. 2014 Apr 16;28(6):935–948. doi: 10.1210/me.2013-1339

Figure 6.

Figure 6.

SGK3 promotes the G1 to S phase cell cycle progression of LNCaP cells through up-regulation of cyclin D1. A and B, LNCaP cells were transfected with the siRNA negative control (siNC) or pooled SGK3 siRNA duplexes (siSGK3) for 4 days. The transfected cells were imaged under a microscope (A) or harvested for Western blotting (B) to determine PARP cleavage and cleaved caspase 3 levels. Scale bar corresponds to 50 μm; ×100 magnification. For Western blotting analysis of apoptosis markers, we also included a positive control, which are LNCaP cells treated with 20 μM proteasome inhibitor MG132 for 24 hours. C, LNCaP cells were transfected with siRNA negative control (LNCaP/siNC) or pooled SGK3 siRNA duplexes (LNCaP/siSGK3). Seventy-two hours after transfection, cells were harvested, fixed in 70% ethanol, and stained with propidium iodide and analyzed by flow cytometry. The percentage of DNA content in each phase was analyzed with ModFit LT 2.0 software. The representative result of 3 experiments was shown. D, Quantitative result of the percentage of DNA content in each cell cycle phase from 3 experiments. The data are expressed as means ± SD. *, P < .05; **, P < .01, by the Student t test. E and F, LNCaP cells were transfected with the siRNA negative control, pooled (E) or 3 individual (F) SGK3 siRNA duplexes, respectively. At 72 hours after transfection, the cells were harvested for Western blotting analysis with the relevant antibodies.

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