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. Author manuscript; available in PMC: 2016 Mar 29.
Published in final edited form as: Nat Cell Biol. 2009 Mar 15;11(4):501–508. doi: 10.1038/ncb1858

Figure 1.

Figure 1

Depletion of rpS6 leads to p53-dependent cell-cycle arrest. (a) Expression kinetics of p21 mRNA after partial hepatectomy in the livers of rpS6fl/flMxCre− (filled diamonds) and rpS6fl/flMxCre+ (open squares) mice, as measured by hybridization of DNA microarrays. Results are shown as means ± s.e.m. for three independent experiments. (b) Protein levels of p53 in the livers of rpS6fl/flMxCre− and rpS6fl/flMxCre+ mice at 30 h after partial hepatectomy were measured with an ELISA kit (see Methods). For each genotype the measurements from two different animals are shown. (c) Northern blot (upper panel) and western blot (lower panel) showing the levels of rpS6 mRNA and rpS6 protein, respectively, in non-transfected A549 cells (lane 1), or A549 cells transfected for 24 h with the indicated siRNAs. (d) Polysome profiles from extracts of A549 cells treated for 24 h with either NSsiRNA (grey line) or rpS6 siRNA2 (black line). The positions of 40S and 60S native subunits and 80S monosomes are indicated. (e) Western blots showing the levels of rpS6, p53 and p21 proteins in cells transfected for 24 h with NSsiRNA or rpS6 siRNA2 in combination with either NSsiRNA or p53 siRNA. (f) Quantification of BrdU incorporation in A549 cells transfected for 24 h with the indicated siRNAs. For each treatment, the values are normalized to values for cells transfected with NSsiRNA. Results are shown as means ± s.e.m. for three independent experiments. Uncropped images of blots are shown in Supplementary Information, Fig. S5.