Xenbase Image ID: 137092

Figure 7. USP15 impacts osteoblastic differentiation in C2C12 myoblasts and modulates BMP signalling in Xenopus embryogenesis. (a) Mouse myoblast cell line C2C12 were transfected with siRNAs targeting mouse FoxO4 or USP15. Cells were serum-starved overnight and treated with or without BMP for 1 h prior to lysis. Extracts were resolved by SDS-PAGE and immunoblotted with antibodies against USP15, pSMAD1, total SMAD1 and GAPDH. (b) C2C12 cells transfected with mouse siFoxO4 or mouse siUSP15 were grown for up to 4 days in the presence of BMP. Cells were lysed and the alkaline phosphatase activity measured using a fluorescence plate reader. Data are represented as mean of three biological replicates and error bars indicate s.d. Representative extracts were resolved by SDS-PAGE and subjected to immunoblotting with antibodies against USP15 and GAPDH. (c) Xenopus embryos were injected with 80 ng of either xUSP15- (xUSP15-MO) or control-MO morpholinos at the one-cell stage and then collected at the indicated stages. Lysates were resolved by SDS-PAGE and immunoblotted with antibodies against pSMAD1 and α-tubulin. (d) qRT-PCR analysis of xVENT1 mRNA expression. Embryos were injected with 80 ng of either USP15-MO or control-MO at the one-cell stage and then animal caps were cut at stage 8.5. The animal caps were collected at the equivalent embryo stage of 10.5 and processed for qRT-PCR. Image published in: Herhaus L et al. (2014) . 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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