Figure 4
PGE2 effects are mediated through specific PGE2 receptor subtypes. (A) hBMSC were cultured in growth medium (GM) or osteogenic medium (OM), and PGE2 receptor gene expression levels determined at day 7 and 14 using RT-qPCR. *p < 0.05, **p < 0.01, ***p < 0.001 as compared to GM using Student’s t-test. (B) RT-qPCR was used to assess the short-term effects (48 h) of siRNA specific for PTGER2 (siPTGER2), PTGER4 (siPTGER4) or both PTGER2 and PTGER4 (siPTGER2/4) on PTGER2 and PTGER4 gene expression in hBMSCs undergoing osteogenesis. *p < 0.05, **p < 0.01, ***p < 0.001 as compared to cells treated with scrambled control siRNA (siControl) using ANOVA. (C) RT-qPCR was used to assess the long-term effects (3, 7 and 10 days) of siRNA specific for both PTGER2 and PTGER4 (siPTGER2/4) on PTGER2 and PTGER4 gene expression in hBMSCs undergoing osteogenesis. *p < 0.001 as compared to cells treated with scrambled control siRNA (siControl) using Student’s t-test. (D) The effects of continuous PGE2 (10 nM) treatment on matrix mineralization in siRNA-treated hBMSCs was assessed at day 15 by Alizarin Red S staining. *p < 0.01, **p < 0.001, as compared to untreated hBMSCs (−PGE2) using Student’s t-test. The data represent triplicate determinations and were replicated at least two times. All values are presented as mean ± S.D.
