Figure 4.

Nuclear and cytoplasmic fractionation of TL-1 cells with and without calcitriol treatment. The TL-1 cell line was treated with 10 nM calcitriol, or appropriate negative controls for 24 hours. Subcellular fractionation was performed to isolate cytoplasmic and nuclear proteins. Total protein loaded per sample was 25 μg, with HSP90 and histone H3 used as markers of cytoplasm and nucleus, respectively. Two identical separate blots were used to probe for all targets. The subcellular fractions were probed for (A) VDR and STAT1 (pY701 and total) and (B) STAT3 (pY705, total). These are representative western blots of three independent experiments.