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. 2012 Nov 1;23(21):4242–4255. doi: 10.1091/mbc.E12-06-0486

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© 2012 Sellin et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 2: — Partitioning of individual septins between soluble and insoluble fractions of cells permeabilized at graded ionic strengths. (A) K562 cells were permeabilized by saponin (0.2%) in either 1× PEM or 0.25× PEM buffer. The released (supernatant [S]) and cell-associated (pellet [P]) proteins were analyzed by Western blot using the indicated antibodies. The release of a cytosolic protein (Op18) and retention of the intermediate filament protein vimentin served as a marker for cell permeabilization. (B) The fraction of the indicated septins within cell pellets after saponin (0.2%) permeabilization with the indicated PEM buffer concentration was determined by quantification of Western blots. SEPT9^total represents all three endogenous isoforms, which appeared indistinguishable with respect to their partitioning. Bar charts represent the mean ± errors of data from duplicate experiments.