Fig. 2. Human LEM2 is a lamina protein of the INM. HeLa cells stably expressing V5-tagged hLEM2 were processed for immunofluorescence using anti-V5 (green) and anti-LAP2 (red) antibodies. DNA was stained with Hoechst (blue). Cells were either extracted with (A) Triton X-100 or (B) digitonin after fixation. Arrows point to mitotic cells, which serve as positive control for antibody reactivity. Bars, 10 µm. (C) Subcellular fractionation of lysates of stable HeLa clones expressing V5-tagged hLEM2. Nuclei were extracted with Triton X-100 and/or salt, or with 7M urea, and soluble supernatant and insoluble pellet fractions were subjected to western blot analysis using anti-V5 and anti-LAP2ß antibodies. Molecular weights in kDa are indicated on the left.