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. 2009 May 8;106(20):8284–8289. doi: 10.1073/pnas.0900641106

Fig. 2.

Fig. 2.

Quantification of the DE-Cad expression levels in wild type, DE-Cad(rescue), and DE-Cad::GFP. (A) A sample Western blot showing the DE-Cad expression levels in wild type, DE-Cad::GFP (see Fig. 2 A and B), and DE-Cad(rescue) homozygous embryos. Embryos were in mixed stages (24-h collection under 25 °C). (Top) Because the majority of DE-Cad proteins undergoes internal cleavage (25), the rat anti-DE-Cad monoclonal antibody (DCAD2) (25) recognizes a single 150kDa band in all 3 samples. (Middle) To confirm the identity of DE-Cad::GFP, we also blotted the same samples with anti-GFP antibody. Only in DE-Cad::GFP sample the antibody recognized a single band around 100 kDa, which corresponds to the carboxyl-terminal half of cleaved DE-Cad::GFP (26). (Bottom) α-tublulin was used as loading controls. (B) Quantitative measurements of DE-Cad protein levels in wild type, DE-Cad(rescue), and DE-Cad::GFP that were based on multiple Western blot results (DE-Cad(rescue): n = 3; DE-Cad::GFP: n = 4).