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. 2016 Oct 18;15(23):3278–3295. doi: 10.1080/15384101.2016.1243189

Figure 3.

Figure 3.

Stable silencing of CycG2 expression attenuates the cell cycle arrest response of MCF7 cells to estrogen withdrawal. (A) Immunoblot of endogenous CycG2 in MCF7 clones expressing CCNG2-targeting shRNAs (1-B and ID3) or a non-silencing control shRNA (NSC) cultured with (+) or without (-) estrogen depletion (DM). Numbers underneath shRNA denominators refer to clone number for the respective stably transfected cell line. (B) One-way ANOVA statistical analysis of CycG2 expression data from > 3 replicate experiments quantified by immunoblotting as in A. (C) Histogram overlays of DNA content in MCF7 control and CycG2 KD clones cultured in E2-depleted (black line, DM days) or normal media (gray area, NT). (D) One-way ANOVA statistical analysis of G1- and S-phase cell cycle distribution of single parameter DNA flow cytometry data analyses similar to that shown in C. (E) Two-parameter flow cytometry analysis of BrdU-labeled DNA in E2-depleted (DM) compared to non-treated (NT) CycG2 KD clones and controls. (F) One-way ANOVA analysis of BrdU incorporation and cell cycle distribution data as shown in E. ***p = < 0.001; **p = < 0.01; *p = < 0.05.