Fig. 4
Role of LATS1 in trophoblast giant cell differentiation. A Western blot analysis of LATS1 and pLATS1Thr1079 in TSCs (Stem cells) and differentiated trophoblast cells. GAPDH was used as an internal control. B Densitometric quantification of the protein bands from A. The level of pLATS1Thr1079 was quantified relative to the basal level of LATS1. C Western blotting showing the coimmunoprecipitation of endogenous LATS1 with LIMK2 in TSCs and differentiated trophoblast cells. Immunoprecipitation (IP) with anti-LATS1 antibody was followed by immunoblot with anti-LIMK2 antibody. Immunoprecipitation using an isotype matched IgG was used to ensure the specificity of the capturing antibody. D Immunoprecipitation demonstrates that LATS1 exists as a protein complex pulled down by anti-LIMK2 antibody. E Western blot analysis showing the probing of LIMK2, pLIMK2Thr505, COFILIN, pCOFILINSer3 and CHRONOPHIN in TSCs and differentiated trophoblast cells. D Densitometric analysis of the protein bands in E using GAPDH as an endogenous control. LIMK2, COFILIN and CHRONOPHIN are normalized relative to the level of the housekeeping gene, whereas the level of pLIMK2Thr505 and pCOFILINSer3 was normalized relative to the basal level of LIMK2 and COFILIN, respectively. Values are represented as mean ± SEM from three independent biological replicates. Statistical analysis was performed using Student’s unpaired t-test, *p < 0.05; **p < 0.005; ***p < 0.0005, ns, nonsignificant