Figure 5
Testosterone-induced down-regulation of Cx43 in granulosa cells was reversed by treatment with 1,25D 3 . (A) Granulosa cells were treated for 24 h with vehicle, testosterone (0.1 or 1 μg/mL) or testosterone (1 μg/mL) plus 1,25D3 (0.1 μM) and fixed in cold paraformaldehyde solution. Cell nuclei were stained with DAPI (left column), and Cx43 levels (green) were examined by confocal microscopy (middle column). The overlay is shown in the right column. (B, C) Granulosa cells were treated with 0.01, 0.1, or 1 μg/mL testosterone for 24 h in the presence of vehicle control or 0.1 μM 1,25D3. The levels of Cx43 protein and Cx43 mRNA were examined by western blot and qRT-PCR analysis, respectively. The results are expressed as the means ± SEM (n = 4–6 in each group). **, p < 0.005 for comparison with the vehicle-treated group. #, p < 0.05 for comparison of two drug groups. The values listed within column indicate the means. Scale bars, 10 μm.