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. 2009 Jun 9;122(13):2311–2321. doi: 10.1242/jcs.046557

Fig. 4.

Fig. 4.

A minor role for p110δ in mESC self-renewal. (A) mESCs were treated with a p110δ inhibitor, IC87114, for 4 days and self-renewal assessed by staining for alkaline-phosphatase activity. The average percentage of alkaline-phosphatase-positive, self-renewing colonies in each condition are shown with s.e.m. (n=4); *P<0.05 following a Student's t-test. The number of total colonies formed in each condition did not vary significantly. (B) Quantitative RT-PCR was conducted to detect knockdown of Pik3cd RNA following transfection with Pik3cd-targeting siRNA or with non-targeting siRNA. Representative data is shown with s.d. and the percentage of knockdowns, compared with control, are indicated. (C) Following transfection, cells were plated for 4 days and self-renewal assessed by alkaline-phosphatase staining. The average percentage of alkaline-phosphatase-positive, self-renewing colonies in each condition are shown with s.e.m. The number of total colonies formed in each condition did not vary significantly. (D) Expression of Nanog was assessed by quantitative RT-PCR following knockdown of Pik3cd. Representative data is shown with s.d. (E) Immunoblotting was carried out to detect p85 protein levels, with SHP-2 reprobing used to confirm equal loading.