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Figure 5. ZC4H2 is required for BMP induced osteogenic differentiation in C2C12 cells. (a) ZC4H2 mRNA level in C2C12 cells with different siRNAs transfected during BMP-induced osteogenic differentiation. C2C12 cells were transfected with negative control or three independent siRNAs targeted for ZC4H2 and then were treated with 50 ng ml−1 rhBMP2 for 3 days to induce osteogenic differentiation. Then total RNAs were isolated and real-time PCRs were carried out. (b–e) ZC4H2 knockdown decreased the expression levels of BMP induced osteogenic markers ALP (b), Osteocalcin (c), Osterix (d) and Type I Collagen (e). The experiments were carried out as described above for panel (a). (f,g) ZC4H2 knockdown reduced rhBMP2-induced ALP activity in C2C12 cells. C2C12 cells were treated for osteogenesis as described above in panel (a) after the indicated siRNAs were transfected. After induction, ALP activity was analysed by an ALP fluorometric kit (f) or NBT/BCIP staining (g). NC, negative control. n.s., not significant; **p < 0.01. Image published in: Ma P et al. (2017) © 2017 The Authors. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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