FIG. 5.

Inflammatory cytokines repress GSK3 inhibitor-induced HSC expansion independent of Akt-mTor signaling. (A, B) BM MNCs were cultured for 2 days in stem cell medium at the indicated CHIR concentrations. DMSO-treated cells were used as a control. Protein lysates were collected for western blotting for p-S6 (A). LSK HSPCs were sorted and stained with p-S6 antibodies. mTor activity was examined by FACS for p-S6 (B). (C, D) BM MNCs were cultured for 9 days in stem cell medium with 0.5 μM CHIR99021 plus indicated concentrations of rapamycin with medium change every other day. The percentages and numbers of LSK HSPCs and LSK-CD150⁺48⁻ HSCs were examined by FACS (C) and functional HSCs were evaluated by CHR assay (A). *P < 0.05. Color images are available online.