Figure 6.

Nuclear trafficking of VDR in presence of geldanamycin (a) or leptomycin B (b). HL60 cells were exposed to 10 nM 1,25(OH)₂D for 3 h. One sample was pretreated with 1 μM geldanamycin (geld) for 1 h before exposure to 1,25(OH)₂D (a). Another sample was treated with 5 mg/ml of leptomycin B (lept B) for the last two hours of incubation (b). After incubation the cells were lysed and fractionated into the cytoplasmic (C) and nuclear (N) fractions. The lysates from equal numbers of cells were separated in SDS-PAGE and blotted to the membrane. The membrane was probed with anti-VDR. Actin was probed as a control of equal loading and transfer of proteins.