Analysis of cell proliferation over a 5-day period of induction with doxycycline (Dox).

<p>(<b>A</b>) Proliferation rate of cells expressing the shRNA targeting FTSJ3. (<b>B</b>) Proliferation rate of cells expressing the scrambled control SC RNA. (The graphs represent one of four independent assays performed using four technical replicates for each cell treatment). Induction of the shRNA targeting FTSJ3 leads to a reduction in cell proliferation. (<b>C</b>) Cell cycle analysis by fluorescence-activated cell sorting (FACS). FACS analyses were performed with HEK293 Flp-In T-Rex cells stably transfected with the pFRT-U6tetO plasmid expressing either a shRNA targeting FTSJ3 (shRNA) or a scrambled control (SC) and induced (+) or not (−) with 5 µg/ml of doxycycline for 5 days. (<b>D</b>) Relative growth rate differences between cells expressing the scrambled control scRNA (sc+/sc−) and cell expressing the shRNA targeting the FTSJ3 mRNA (sh+/sh−) induced (+) or not (−) with doxycycline for the 120 h time point. (<b>E</b>) Immunoblot analysis showing the levels of the FTSJ3 and p53 proteins in cells expressing either the shRNA targeting the FTSJ3 mRNA or the scrambled control induced (+) or not (−) with doxycycline. GAPDH was used as a gel loading control. The levels of FTSJ3 are reduced as expected and the levels of p53 are not affected.