Activation of MAP kinase and AKT signaling pathways in Lemd2-deficent embryos.

(A, B, D-G) Western blot analysis of protein extracts from Lemd2^+/+ and Lemd2^Gt/Gt embryos at E10.5. Samples from 4 separate embryos of each genotype are shown. All extracts were from embryos with beating hearts. (A) Labeling with anti-Lem2 and anti-GAPDH. (B, D-G) Labeling with antibodies to the phosphorylated activation sites of various kinases or kinase effectors (upper panels) and to nonphosphorylated epitopes of the corresponding proteins (lower panels). (B) Anti-ERK1/2. (D) Anti-p38. (E) Anti-JNK1/2. (F) Anti-AKT. (G) Anti-Smad2. Error bars indicate standard deviations (*p < 0.0005, **p < 0.005, ***p < 0.05, ns “not significant”). (C) Immunofluorescence staining of sagittal cryosections of Lemd2^+/+ and Lemd2^Gt/Gt hearts and somites at E10.5 with anti-phospho-ERK1/2 antibodies. All images were captured and displayed with exactly the same parameters.