Fig. 7
Inhibition of LIMK2 impedes TGC formation. A Western blot analysis of pLIMK2Thr505, COFILIN, pCOFILINSer3 in trophoblast cells treated with the indicated doses of BMS-3. Control cells received treatment with equivalent amount of vehicle (DMSO). Cells were harvested after 72 h of treatment. B Densitometry-based quantification of the protein bands from A using ImageJ. Band intensities of COFILIN were normalized using GAPDH as endogenous control. The level of pLIMK2Thr505 and pCOFILINSer3 was normalized relative to the basal level of the respective proteins. C Quantitative real-time PCR analysis of Prl2c2 transcript in trophoblast cells treated with 5 µM and 10 µM BMS-3. D Representative confocal photomicrographs of trophoblast cells under similar experimental conditions. Counterstaining of the nuclei was done using Hoechst (blue). DyLight™ 554 Phalloidin (red) was used to stain the F-actin filaments. Disintegration of the actin filaments has been represented in the boxed areas which have been magnified in the rightmost panel. Scale bar: 10 µm. Magnification: × 72 D ImageJ-based quantification of the nuclear surface area/cell of trophoblast cells upon LIMK2 inhibition. Error bars represent the SEM of 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