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Figure 2


Fig. 2. Characterization of SNX9, SNX18 and SNX30 proteins in cells. (A) HeLa-cell cytosol was immunoprecipitated with antisera against the three paralogs, and with pre-immune serum, and analyzed with SDS-PAGE. The gel was stained with Coomassie. Bands corresponding to IgG and to immunoprecipitated SNX9, SNX18 and SNX30 are indicated. The identity of SNX18 and SNX30 was determined by MALDI-TOF. Arrows show the positions of dynamin 2 (upper) and aldolase (lower). Inset below shows an immunoblotting analysis with respective antibody of immunoprecipitates from a total HeLa-cell detergent extract. Note that individual paralogs do not form complexes with other family members. (B) Epifluorescent micrograph of a HeLa cell transfected with myc-SNX30 and stained with mouse anti-myc, rabbit anti-SNX9 and chicken anti-SNX18. Scale bar: 10 µm. Enlarged pictures show no overlap in localization of the three proteins. (C) A membrane fraction from HeLa cells was separated by density equilibrium centrifugation, and collected fractions were analyzed for density (triangles) and total protein amount (circles). The distribution of the indicated proteins was analyzed by immunoblotting.





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