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Figure 3. A: Control of specificity of the anti-XMRF4 antibody: One nanogram of synthetic Flag-tagged XMRF4-a (1) or -b (2) mRNA was bilaterally injected into the animal pole of two-cell stage embryos. Ten micrograms of egg homogenates was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western blotting. The affinity-purified anti-XMRF4 antibody recognizes the two XMRF4 proteins in the same way as the anti-Flag antibody. No band was obtained in uninjected embryos (data not shown). B: Whole mount immunostaining using an affinity-purified polyclonal antibody against Xenopus MRF4 at stage 24 (up), 26 (middle), 28 (down). The anterior side of the embryos is on the left. Arrowhead, posterior unsegmented mesoderm. C: Indirect immunofluorescent detection of Anti-XMRF4 (green) and 4 prime ,6-diamidine-2-phenylidole-dihydrochloride (DAPI) staining (blue) on longitudinal cryosection of somitic muscles at stage 28. No labels were detected with secondary antibody alone (data not shown). |
