Fig. 5
Effect of RAB14 on cellular migration and invasion. (A) Changes in levels of RAB14 protein in TE7, FLO-1, and SK-GT-4 cells following transfection of 80 pM RAB14-siRNA. The blots in this figure were cut prior to hybridization with antibody to RAB14 and GAPDH. Relative signal intensity was calculated as explained in Fig. 1E. (B) Migration and invasion assays in real time. Following silencing of RAB14 in TE7, FLO-1, and SK-GT-4 cells as described in (A), migration (a) and invasion (b) assays using XCELLigence-RTCA system were performed. Representative bar diagrams for three biological and technical replicates are shown. * denotes statistical significance (p < 0.05). (C) Changes in levels of RAB14 in hESO cells following transfection of 2.5 μg of a RAB14 expression plasmid. The blot in this figure was cut prior to hybridization with antibody to RAB14 and GAPDH. (D) Migration (a) and invasion (b) assays in real time using XCELLigence RTCA system were performed following overexpression of RAB14 in hESO cells as described in (C). Representative bar diagrams are shown. * denotes statistical significance (p < 0.05). (E) Following overexpression of pre-miR-214-3p (50 nM) (middle lane) or pre-miR-214-3p followed by 2.5 μg of RAB14 expression plasmid (last lane), RAB14 levels were measured by immunoblot and compared to control miR transfection. Relative signal intensity was calculated as explained in Fig. 1E. The blot in this figure was cut prior to hybridization with antibody to RAB14 and GAPDH. (F) Migration (a) and invasion (b) assays were performed in the samples explained in E. Mean ± S.D. is shown and statistical significance is indicated by * (p < 0.05)