Figure 6.

LIF mediates the effects of PKIγ knockdown on osteogenesis and adipogenesis in hMSCs. hMSCs reverse transfected with non-targeting siRNA duplexes as a control (C) or siRNA targeting PKIγ (KD) were incubated for 6 days in osteogenic medium (A–E) or 3 cycles of adipogenic induction medium (3 days) and adipogenic maintenance medium (2 days) (F–G). Culture medium was supplemented with 10 ug/ml of either control immunoglobulin (IgG2B) or the anti-human LIF monoclonal antibody (LIF MAb). mRNAs encoding members of the PKI family (A–C), osteoblastic marker genes (D–E), and adipogenic marker genes (F–G) were measured by quantitative real-time RT-PCR. C/EBPA and leptin mRNA were undetectable in all groups with adipogenic media (not shown). * denotes p < 0.05 and NS denotes not significant. Horizontal bars represent the means of the 3 independent experiments indicated by the individual symbols that represent means of 3 culture wells/group, each assayed in triplicate.