Reduced proliferation but normal osteogenic differentiation of bone marrow stromal cells from Akt1 deficient male mice.

<p><b>A</b>. Experimental scheme: bone marrow stromal cells from 20-week old 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 10 days. <b>B</b>. Measurement of cell number by daily counting (mean ± S.D., n = 3 experiments; *- p<0.01, **- p<0.003 vs. Akt1 WT). <b>C</b>. Measurement of gene expression by RT-PCR on days 5, 7, and 10 after incubation in OM plus BMP2 for Akt1, Akt2, Runx2, osteocalcin (OCN), and S17 mRNAs. <b>D</b>. Immunoblots of whole cell protein lysates on days 7 and 10 after incubation in OM plus BMP2 for Akt1, Akt2, total Akt, phospho (p) Akt, Runx2, and α-tubulin. <b>E</b>. Alkaline phosphatase (AP) and Alizarin red staining on day 10.