Figure 2. Knockdown of endogenous GSK3β by siRNA inhibits osteoblast differentiation in ADSCs.

(A–C) ADSCs were transiently transfected with 10 nM GSK3β siRNA or a nonrelevant siRNA (scramble). After 48 hours, GSK3β silencing was determined by both mRNA (upper, RT-PCR analysis) and protein (lower, immunoblot analysis) levels, respectively. GAPDH is provided as a loading control (A). GSK3β silencing reduced ALP, osterix, and osteopontin mRNA expression determined by real time PCR analysis (B). Silencing GSK3β abolished ALP activity and matrix mineralization as determined by ARS (C). Data represent mean ± S.D. and are representative of at least 3 experiments. *p≤0.01, significantly different from scramble siRNA transfected group.