Fig. 3
RNF144B promotes ovarian cancer cell proliferation, colony formation, and migration. Cell viability assays were performed on (a) PEO4 and (b) OVCAR3 cells stably overexpressing RNF144B compared to mock and parental cells. The number of viable cells was measured using an MTT reagent and spectrophotometric absorbance was taken at 570 nm, p = 0.0003 & 0.04. Ovarian cancer cells (n = 1000) (c) PEO4 (clone-3) and (d) OVCAR3 (clone-1) stably expressing RNF144B were plated in 6-well plates. Cells transfected with empty vector pCDNA4 were served as controls. After two weeks, the colonies formed were stained and counted using the grid method, p = 0.001 and < 0.0001. A wound healing assay was performed in (e) PEO4 (clone-3) and (f) OVCAR3 (clone-1) cells stably expressing RNF144B. Wound closure was captured at 0, 24, and 48 h at 4 × magnification. Wound closure was calculated by measuring the distance covered by the cells relative to the initial wound area determined at 0 h using image J software. The wound closure data is expressed in percentages, p = 0.0002 & 0.001. A transwell Matrigel invasion assay was performed on (g) PEO4 (clone-3) cells expressing RNF144B or empty vector. Cells were added to the upper chamber of the transwell insert. The cells that invaded through the Matrigel were fixed and stained. The average number of migrated cells was compared between RNF144B transfected and control, p = 0.09. All experiments were performed in triplicate, with three independent replicates for each. Statistical significance was calculated using the two-tailed student’s t-test, and the results are represented as mean ± SD.
