Fig. 3

RT-PCR and immunocytochemistry analysis show distinctive mRNA and protein expression in Weri-Rb-1 and Y79 cells. A-B. ELTD1 (A) and GPR125 (B) mRNA expression in Y79 and Weri-Rb-1 cell lines. Results expressed as relative fold change of target gene over HPRT1 as control. C-J. Low magnification images of anti-ELTD1 (green) antibody labeling show punctate clusters in cell to cell adhesion sites mostly prominent in the more adhesive Weri-Rb-1 cells. C1-J1. High magnification images of insets in panels F and J emphasizing the highly intense ELTD1 clusters in cell to cell adhesion sites (white arrows; Weri-Rb-1 cells), cytoplasm and nucleus of both Rb cell lines. K-R. Low magnification images of anti-GPR125 (green) antibody labeling show uniform labeling in the nucleus and cytoplasm of the less invasive Weri-Rb1 cells, while in the more invasive Y79 cells it showed enriched labeling in presumptive leading edge sites (white arrowheads). K1-R1. High magnification images of insets in panels N and R emphasizing the highly intense GPR125 labeling in presumptive leading edges of Y79 cells. D, H, D1, H1, L, P, L1, P1. Anti-Texas-Red conjugated Phalloidin antibody labeling of F-actin fibers in the Weri-Rb1 and Y79 cells. E, I, E1, I1, M, Q, M1, Q1. DAPI labeled nuclei of Weri-Rb-1 and Y79 cells. F, J, F1, J1, N, R, N1, R1. Merged images which were equally adjusted to improve representation. Scale bar for panels C-J and K-R is 10 μm and for panels C1-J1 and K1-R1 is 5 μm