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.