Fig. 6
Association between overexpression of Nodal and cell proliferation in endometriosis, OCCCa, and OEmCa. a Two independent stable Nodal-expressing ES-NoD cell lines and mock cells were seeded at low density. The cell numbers are presented as mean ± SD. P0, P2, P4, P6, and P9 indicate 0, 2, 4, 6, and 9 days after cell passage, respectively. b Western blot analysis for the indicated proteins in the stable Nodal-expressing ES-NoD and mock cells for the times shown following restimulation with 10% serum after serum starvation for 6 h. c Ishikawa cells were transfected with p21waf1, p27kip1, and Cyclin A2 reporter constructs, together with cotransfection of Smad2 and Nodal. Relative activity was determined based on arbitrary light units of luciferase activity normalized to pRL-TK activity. The activities of the reporter plus the effector relative to that of the reporter plus empty vector are shown as means ± SDs. The experiment was performed in duplicate. d Left: staining by hematoxylin and eosin (HE) and IHC for Nodal and Ki-67 in endometriosis, OCCCa, and OEmCa. Original magnification, × 200. Middle and right: Ki-67 labeling indices in endometriosis-OCCCa (middle) and endometriosis-OEmCa (right). The data shown are means ± SDs. E1, endometriosis without carcinoma; E2, endometriosis with carcinoma; Ca, carcinoma. e Ki-67 labeling indices in OCCCa (left) and OEmCa (right) between Nodal positive (P) (score ≧ 1) and negative (N) categories (score = 0). The data shown are means ± SDs