FIG. 1.

Evaluation of vitamin D receptors in mouse embryoid bodies. Embryoid bodies were cultured with control medium or medium supplemented with BGP. Real-time qPCR was performed on samples at embryoid body formation (day 0), or after 7 or 14 days in culture. mRNA expression of Vdr (A) and Pdia3 (B) were measured and are presented as normalized to GAPDH. *P<0.05, versus day 0; ^#P<0.05, versus control. VDR and Pdia3 proteins were analyzed by western blot using GAPDH as a reference (C). Ratios of VDR/GAPDH (D) and PDIA3/GAPDH (E) were calculated using pixel intensity of western blot bands. *P<0.05, versus Day 0. Immunofluorescent staining of embryoid bodies was performed for VDR and Pdia3 after 14 days culture in control medium or medium supplemented with BGP (F). Cells were stained for β-actin (red), DAPI nuclear staining (blue), and antibodies against either VDR or Pdia3 (green) and imaged at 63×or 100×magnification (scale bar-50 μm). BGP, β-glycerophosphate; qPCR, quantitative polymerase chain reaction; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; VDR, vitamin D receptors; PDIA3, protein disulfide isomerase family A, member 3; DAPI, 4′,6-diamidino-2-phenylindole. Color images available online at www.liebertonline.com/scd