Figure 2.

Catalytically active D3 is present in the cell nucleus. A, Immunofluorescence of normoxic SK-N-AS cells stained with α-D3(CT) antibody. B, Same as in A except that α-nuclear lamin antibody was also used for staining and images were overlaid. C, Western blotting with α-D3(NT) antibody or α-BiP antibody. SK-N-AS cells homogenized with the ball homogenizer (Isobiotec). Nuclear pellet was obtained after centrifugation at 800 × g and washing. Supernatant was centrifuged at 200,000 × g to yield the membrane pellet. D, D3 activity in nuclear or membrane fractions shown in C; data are the mean ± SEM; n = 2 per group; experiments were repeated 3 times.