Xenbase Image ID: 78198

Fig. 6. Hypoxia does not affect somitogenesis but affects the accumulation of specific proteins in muscle cells. (A-D) Stage 25, control (A-C) and 5% O2-exposed embryos (B-D). Sections were nuclei stained with Hoechst (blue) and immunostained with the F59 antibodies (red; A-B) or with the 12/101 antibodies (red; C-D). Both stainings disappeared in hypoxic embryos. (E) Western blot analysis using the 12/101 antibodies and anti-alpha tubulin antibodies as a loading control. The 12/101 protein level was weaker in embryos exposed to hypoxia. (F-Q) Stage 25 control embryo and 5% O2-exposed embryos sectioned and stained with Hoechst (blue) and immunostained with the anti-MyoD (G, J) and anti-MRF4 (M,P) antibodies (red). No significant differences in the localisation and expression of MyoD and MRF4 were detected. (R-S) qRT-PCR of Myh 1, 4 and 8 mRNAs (R) and MyoD mRNA (S) in normoxia (21% O2) and hypoxia (10 and 5% O2) at stage 25. The amount of mRNAs remained constant regardless of oxygen availability. (T-W) Stage 25, 21% and 5% O2-exposed embryos, sectioned and stained with Hoechst (blue) and immunostained with the anti-P63 (red; T,U) and anti-acetylated tubulin antibodies (red; V, W). P63 was still expressed in nuclei of cells and acetylated tubulin in ciliated cells of the epidermis. Scale bar = 100 μm. Image published in: Hidalgo M et al. (2012) Copyright © 2012. Image reproduced with permission of the Publisher, The Company of Biologists Ltd. Larger Image Printer Friendly View

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