FIG. 1.
Detection of human Pit2 by SDS-PAGE and Western blotting. CHO (−) and CHO-Pit2-V (+) cells were grown in the presence of 1 mM Pi. Cell extracts were prepared in 1% Triton X-100-NH buffer and analyzed by Western blotting using a rabbit anti-human Pit2 serum. For the supernatants, 5% of total cell extract volume or of the volume of supernatants from a centrifugation at 13,000 × g for 30 min or at 100,000 × g for 1 h was analyzed. For the pellets, all material pelleted from cell extracts was analyzed. Samples were not heated before loading on the gel. Pit2 species are labeled a, b, and c. Molecular mass markers are in kilodaltons.