Figure 3.

Effects of prednisolone and vitamin D₃ administration on the level of glucocorticoid receptors in rat bone marrow (BM): 1—control; 2—prednisolone administration (5 mg/kg of b.w.); and 3—prednisolone and vitamin D₃ (1,000 IU/kg of b.w.) administration. Proteins from total BM lysates were separated by SDS-PAGE and western blot analysis was performed using the antibodies against GR. Representative immunoblot and quantification of three experiments (A) are shown. Protein levels were normalized to β-actin. The bar graphs of GR are presented as means ± SEM (n = 6/group). GR-positive BM cells: representative histograms (count—the number of events; FL1 LOG—fluorescence intensity) and quantification of GR-positive cells documented by flow cytometry analysis (B). All data are shown as means ± SEM; *p < 0.05 vs. control, ^#p < 0.05 vs. prednisolone administration. Confocal microscopy of GR-positive (green fluorescence) BM cells (C) shows a significant decrease in the number of GR-positive cells after prednisolone administration. Hoechst 33342 (blue fluorescence) was used for nuclear staining. Scale bars indicate 50 µm (magnification 40×).