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Fig. 7. Exogenous expression of XMyf5 rescues the phenotype of p38-knockdown embryos. (A) Embryos were injected with XMyf5 MO. DLMZ explants were dissected at stage 10.25 and cultured until sibling embryos reached early neurula stages. (B) Embryos were injected with XMyf5 MO and cultured to tadpole stage. Embryos were fixed and taken for whole-mount in situ hybridization using a probe of muscle actin. 95% of control embryos displayed segmental phenotype (n = 38). 92% of MO-injected embryos lost the segmented phenotype (n = 41). (C) Embryos were injected with the XMyf5 MO. 18 DLMZ explants were dissected at stages 10.25 and cultured until sibling embryos reached early neurula stages (stage 18). RNA was isolated and RT PCR analysis was performed with primers to MHC, desmin, tropomyosin, muscle actin and EF1α serving as a control for quantifying RNA levels in each sample. Control-RT PCR reactions were performed (not shown). (D) Embryos were injected with XMyf5-encoding RNA (0.8 ng), and p38α MO and cultured until control embryos reached stage 29/30. Embryos were fixed and analyzed by whole-mount in situ hybridization using the muscle actin probe (upper panel). Statistical analysis relates to the percentage of embryos displaying normal segmented phenotypes. Embryos were injected as described above. 18 DLMZ explants were dissected at stages 10.25 and cultured until sibling embryos reached early neurula stages (stage 18). RNA was isolated and RT PCR analysis was performed with primers to desmin, tropomyosin, muscle actin and EF1α (lower panel). |
