Fig. 2

Stimulation of tissue factor (Tf) expression by eCG/hCG in ovarian preovulatory follicles. a, The level of tissue factor mRNA was detected in the isolated granulosa (GC) and theca (TC) cells of the preovulatory follicles using real-time PCR. Data are expressed as the mean ± SEM of three experiments. *, P < 0.05 vs. 0 h. b, Total lysates (30 μg protein/lane) extracted from the ovaries were analyzed by western blotting using anti-tissue factor polyclonal antibody (n = 4). Molecular weight is indicated to the left and the size of the tissue factor protein is indicated to the right using arrows. Protein loading was assessed using glyceraldehyde-3-phosphate dehydrogenase (Gapdh). c, Immunofluorescence analysis was performed to determine expression of the tissue factor protein in the preovulatory follicles. Fluorescence was analyzed by confocal microscopy after staining the samples with Alexa Flour 633 fluorescence antibodies (red color). Nuclei were stained with 4′, 6-diamidino-2-phenylindole (DAPI). Data are representative of four independently performed experiments. Arrowhead, Oocyte; asterisk, cumulus cells; arrow, theca cells; POF, preovulatory follicle; GC, granulosa cells; TC, theca cells. Scale bar, 200 μm