PRKD1 regulates HDAC7 localization in immortalized hMSCs. (A) Western blot images of HDAC and PRKD1 phosphorylation levels during osteoblast differentiation, n = 2. (B) Western blot of HDAC and PRKD1 phosphorylation 30 min and 1 h after induction of osteoblastic differentiation, with and without PRKD1 inhibitor (PKDi), n = 2. (C) Detection of HDAC7 in unstimulated hMSCs (Control), 30 min after osteoblastic induction (30 min OD) with and without PKDi or PMA-treated hMSCs, n = 3. (D) mRNA levels of CTGF, CEBPD, CYR61, and ALPL in relative expression 24 h after osteoblastic differentiation with and without PKDi. (E) Quantitation of alkaline phosphatase (ALPL) activity on day 6 of osteoblast differentiation of hMSC transfected with control nontargeting siRNA (siCtrl) or two independent siRNAs targeting PRKD1 (siPRKD1 #1, siPRKD1 #2). For D and E: (*) P < 0.05, Student's t-test; n = 3.
