Fig. 2

ROR2 regulates the expression of cell-cycle markers and delays cell-cycle progression. Western blot analysis of the expression of cell-cycle markers in A375 and UACC903 overexpressing ROR2 (A, E) or M2 and MeWo upon ROR2 silencing (C, F). The graphs show the mean ± S.D. of each protein’s levels in cells transduced with either ROR2 cDNA or shRNA normalized to the corresponding loading control and expressed as the fold change (FC) relative to the corresponding control cell line (Empty or scramble). Statistical significance was tested by a one-tailed Student’s T-Test or ANOVA (MeWo) (n = 3). B, D Assessment of RB localization by immunofluorescence upon ROR2 overexpression (B) or silencing (D). A375-Empty and A375-ROR2 (B) or M2-scramble and M2-shROR2 (D) were stained with DAPI and anti-RB antibody. Graphs show the quantification of RB nuclear/cytosolic localization. Statistical significance was tested by a one-tailed Student’s T-Test (n = 3). G, H Flow cytometry analysis of Cyclin E expression upon ROR2 overexpression (G) or silencing (H). Bar graph shows the mean of Cyclin E MFI (Mean Fluorescence Intensity) ± S.D. Statistical significance was tested by a one-tailed Student’s T-Test (G) or ANOVA (H, MeWo) (n = 3). *: p < 0.01, **: p < 0.001, ***: p < 0.0001, n = 3