Fig. 5. eFM inhibits XmyoD expression in gastrula but not neurula embryos. (A) (i) RNA was extracted at stages 11 and 14 from whole embryos, embryos injected with 80 ng, and embryos injected with 40, 60 or 80 ng of eFM. Total RNA (10 µg) was assayed by RNAase protection for expression of the mesodermal markers XmyoD and Xsna and for the loading control ODC; (ii) RNA was extracted from whole embryos at stage 13, embryos injected with 80 ng of eFM, embryos injected with 200 pg of ssbFGF, and embryos injected with 80 ng of eFM and 200 pg of ssbFGF. Total RNA (10 µg) was analysed by RNAase protection for expression of the marker genes XmyoD, Xbra and the loading control ODC. (B) Embryos injected unilaterally on the left hand side with 40 ng of eFM were assayed at (i) gastrula stage 10.5 and (ii) neurula stage 14 for XmyoD gene expression by in situ hybridisation. (iii) Gastrula stage 10.5 embryos injected unilaterally on the left-hand side with 40 ng of eFM were also assayed by in situ hybridisation for expression of the mesodermal marker Xsna.
