Skip to main content
. 2018 Jun 7;9:303. doi: 10.3389/fendo.2018.00303

Figure 4.

Figure 4

Vitamin D auto/paracrine system in bone marrow (BM) and vitamin D bioavailability after prednisolone and vitamin D3 administration: 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. 25OHD concentration was measured by ELISA (A). Immunoblotting analysis of CYP27B1 in rat BM: the bar graphs of CYP27B1 (B) are presented as means ± SEM (n = 6/group) and representative immunoblots are shown next to the bar chart and quantified using β-actin as a loading control. Quantitative polymerase chain reaction of Vdr in rat BM (C); data were normalized to Gapdh and pooled from two independent experiments (n = 6 rats/group). Quantification of VDR-positive BM cells using flow cytometry analysis (D). Data are shown as means ± SEM; *p < 0.05 vs. control, #p < 0.05 vs. prednisolone administration. Immunocytochemical analysis of double RANK-positive (green fluorescence) and VDR-positive (red fluorescence) BM cells (E). Hoechst 33342 (blue fluorescence) was used for nuclear staining. Scale bars indicate 10 µm (magnification 100×). Red arrows show co-localisation of RANK and VDR in BM mononuclear osteoclast precursors (OCPs) in control group as well as in multinuclear OCPs in prednisolone and prednisolone + vitamin D3 group.