Normal proliferation and differentiation of calvarial osteoblasts from Akt1 deficient mice.

<p><b>A</b>. Experimental scheme: calvarial cells from newborn WT and Akt1 KO mice were incubated in growth medium for 4 days, and then in osteogenic medium (OM) plus 200 ng/ml BMP2 for up to 7 days. <b>B</b>. Measurement of cell number by daily counting. <b>C</b>. Measurement of gene expression by RT-PCR on days 1, 3, 5, and 7 after incubation in OM plus BMP2 for Akt1, Akt2, Runx2, OCN, and S17 mRNAs. <b>D</b>. Immunoblots of whole cell protein lysates on days 7 and 10 after incubation in osteogenic medium (OM) plus BMP2 for Akt1, Akt2, total Akt, pAkt, Runx2, and α-tubulin. <b>E</b>. Alkaline phosphatase (AP) activity on days 5 and 7, and Alizarin red staining on day 7.