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. 2011 Mar 10;2(3):e127. doi: 10.1038/cddis.2011.10

Figure 4.

Figure 4

NPR-A knockdown reduced ES cell propagation, prolonged the G1 phase and upregulated phosphorylated Akt in murine ES cells. (a) Quantification of ES cells 48 h after transfection with control siRNA or NPR-A siRNA (n=4). (b) Effect of NPR-A knockdown on cell-cycle progression, showing the percentages of cells in the G1, S and G2/M phases of the cell cycle. (c) Quantification of cell-cycle phases (n=3). (d) RT-PCR analysis of p21 and cyclin D1 mRNA 48 h after ES cell transfection with control siRNA or NPR-A siRNA. (e) Real-time PCR analysis of ES cells treated as described in panel d. (f) Western blot analysis for p21 48 h after siRNA transfection. (g) Western blot analysis for phosphorylated Akt (Ser473; p-Akt) and total Akt 48 h after siRNA transfection. β-Actin was used as a loading control. (h) Relative levels of the p-Akt protein were quantified after normalization to total Akt. Data represent mean±S.D. (n=3); *P<0.05 or **P<0.01 (two-tailed t-test)