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Xenbase Image ID: 158913
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Figure 2. ZC4H2 regulates BMP signalling. (a,b) The effect of ZC4H2 knockdown or overexpression on the expression of the BMP reporter Id1-Luc in Xenopus animal caps. (a) Schematic drawing showing the experimental strategy for reporter assays in Xenopus animal caps. Embryos were injected with the indicated MO/mRNAs/plasmids in both blastomeres at the two-cell stage and the animal cap tissues were dissected at stage 8 and then were cultured for 20–24 h before harvesting. RLU, relative light units. *p < 0.05. (c–j) ZC4H2 over-expression enhanced while knockdown reduced rhBMP2-induced expression level of Id1-Luc reporter in Hep3B (c–f) and C2C12 cells (g–j). Note that BMP treatment has no effect on either exogenous and endogenous ZC4H2 mRNA levels in both the cell lines (c,e,g,i). Hep3B or C2C12 cells were transfected with the indicated plasmids or siRNAs together with Id1-Luc reporter plasmids and then treated with rhBMP2 for 24 h. Luciferase activity was measured at 48 h after transfection. RLU, relative light units. n.s., not significant. *p < 0.05; **p < 0.01. NC, negative control. (k,l) ZC4H2 knockdown in C2C12 cells decreases mRNA levels of Id1 (k) and Id2 (l) induced by rhBMP2. C2C12 cells were transfected with the indicated siRNAs and treated with rhBMP2 for 24 h before harvesting. Total RNA was extracted for real-time PCR analysis at 72 h after transfection. **p < 0.01. NC, negative control. 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
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