Figure 5.
Overexpression of Smad1 rescues the repression of osteogenesis by miR-203-3p. (A) RT-qPCR analysis of levels of Smad1 expression in C3H10T1/2 cells transfected with Smad1 overexpression plasmid and NC plasmid. (B) Smad1 protein expression in C3H10T1/2 cells transfected with Smad1 overexpression plasmid or NC plasmid and agomir-203-3p or agomir-NC was detected by western blot analysis. (C and D) Smad1 overexpression plasmid or NC plasmid and agomir-203-3p or agomir-NC were co-transfected into C3H10T1/2 cells, followed by osteogenic induction for 7 days. (C) RT-qPCR was used to analyze the expression levels of Runx2, Alp, Osterix and OC. (D) Histochemical detection of ALP in C3H10T1/2 cells (magnification, ×1.4). Overexpression of Smad1 appeared to attenuate the miR-203-3p-mediated suppression of osteogenic differentiation. Values are expressed as the mean ± standard deviation (n=3). ***P<0.001. miR, microRNA; NC, negative control; Agomir, miR agonist; Antagomir, miR antagonist; ALP, alkaline phosphatase; Runx2, runt-related transcription factor 2; OC, osteocalcin; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.
