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. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: Aging Cell. 2011 Aug 24;10(6):962–971. doi: 10.1111/j.1474-9726.2011.00735.x

FIG. 5. Role of CREB signaling in mediating PTH upregulation of CYP27B1/1α-hydroxylase.

FIG. 5

Groups of hMSCs were treated by electroporation with PBS (C, control), with non-silencing control siRNA (NC), or with 100 pmole of CREB-siRNA (CREB-siRNA).

(A) Photomicrographs show cultures of control (C), non-silencing control (NC) and hMSCs transfected with 100 pmole of CREB-siRNA (CREB-siRNA 100 pmole) at 200 × magnification.

(B) Western immunoblot shows p-CREB, CREB, phospho-IGF-IR (p-IGF-IR), total IGF-IR (IGF-IR), CYP27B1, and β-actin protein levels in controls and in transfected cells.

(C) Western immunoblot shows p-CREB, CREB, CYP27B1, p-IGF-IR, IGF-IR, and β-actin in transfected hMSCs after 2 hours or 8 hours in the absence or presence of 100 nM PTH1-34.

(D) Western immunoblot shows p-CREB, CREB, CYP27B1, and β-actin in hMSCs after 2 hours or 8 hours in the absence or presence of 100 nM PTH1-34 ± 30 µM KG-501. Human MSCs were incubated with the indicated concentration of KG-501 for 1 hour before stimulation with PTH1-34. Results are representative of 3 independent experiments.