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Fig. 6. Analysis of endogenous Ldb1 protein levels. (A) XRnf12 downregulates endogenous Ldb1 expression level in a RING-dependent manner. Animal caps were dissected at the blastula stage (stage 8-9) from embryos injected with the mRNAs (2 ng/embryo) indicated, and collected at the gastrula stage (stage 11). Endogenous Ldb1 expression levels were analyzed by western blotting using anti-Ldb1/CLIM2 (N-18) antibody. (B) Ldb1 does not exhibit dorsal-to-ventral (DV) difference in protein expression levels as revealed by western blot analysis. DMZ and VMZ explants dissected at the gastrula stage (stage 11) were compared for Ldb1 expression using anti-Ldb1/CLIM2 (N-18) antibody. (C) Ldb1 is uniformly expressed in the bisected mid gastrula embryo as revealed by immunostaining using anti-Ldb1/NLI antibody. Animal side is to the top, dorsal to the right. Arrowhead indicates blastopore.