FIGURE 1.

Fluorescence microscopy and immunoblotting evidence of FADS localization in BHK-21 cell nuclei. A, fixed and permeabilized BHK-21 cells were incubated with the polyclonal anti-FADS antiserum followed by incubation with an Alexa Fluor 488-conjugated anti-mouse antibody (green). B, nuclei were stained with Hoechst 33658 (blue, B). C, colocalization of FADS with the nuclear marker is depicted in white. Scale bar, 50 μm. D, immunoblotting of different anti-FADS-immunoreactive bands in BHK-21 cells. BHK-21 cells were lysed using 1% Triton X-100 as described under “Experimental Procedures.” Both the Triton X-100-soluble (TX sol) and Triton X-100-insoluble (TX ins) fractions were analyzed by immunoblotting with the anti-FADS antiserum. His₆-hFADS2 (0.03 μg) was also loaded (hFADS2, first lane) as a control. The same PVDF membrane was tested with the anti-β-actin antibody. The arrows indicate the position of the main anti-FADS-immunoreactive bands.