Fig. 3
From: A functional role of LEFTY during progesterone therapy for endometrial carcinoma
Regulation of cell cycle progression by LEFTY and pSmad2 in Em Cas. a Western blot assay for the indicated molecules in the Ishikawa and Hec251 cells, which were synchronized in the G1 phase by treatment with 50 nM rapamycin, in the early S phase with 2 μg/mL aphidicolin, or in the G2/M phase with 0.25 μg/mL nocodazole for 24 h. Asy, asynchronous. b Western blot assay for the indicated molecules in Ishikawa cells treated with 0, 2, and 4 ng/mL LEFTY2 for 24 h (left), and transfection of 50 and 70 nM LEFTY1 siRNA for 72 h (right). c Western blot analysis for the indicated proteins in Ish-shL1#43 and #81 stable cells and the mock cells for the times shown following restimulation with 10% serum after serum starvation for 24 h. d Ishikawa cells were transfected with cyclin A2, cyclin D1, p21waf1, and p27kip1 reporter constructs, in addition to cotransfection with Smad2 or LEFTY1. 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. (E) Left: staining by hematoxylin and eosin (HE) and IHC for LEFTY, cyclin A2, and Ki-67 in Em Cas. Original magnification, ×100. Right: labeling indices for cyclin A2 and Ki-67 in Em Cas. The data shown are means ± SDs