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. 2011 Mar 15;22(6):842–857. doi: 10.1091/mbc.E10-07-0570

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© 2011 Rossi and Brennwald. 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 3: — GFP-Sro7 polarization to sites of active growth is dramatically perturbed in the myo2-iq1,5 mutant. (A) Wild-type strain expressing GFP-Sro7 (CEN) was grown in selective media to midlog at 25°C and then fixed and processed for fluorescence microscopy. Bar, 1 μm. (B) Wild-type and myo2-iq1,5 strains expressing GFP-Sro7 (CEN), GFP-Sec4 (CEN), and GFP-Mlc1 (CEN) were grown in selective media at the permissive temperature (32°C) and then fixed and processed for fluorescence microscopy. Quantitation of polarized tagged proteins in wild-type and myo2-iq1,5 mutant cells were obtained by scoring a minimum of 50 cells at emerging bud sites and bud tips. Bar, 1 μm. (C) Wild-type and myo2-iq1,5 mutant cells were grown overnight at permissive temperature (32°C) and then shifted to 25°C for 8 h before being spheroplasted, lysed, and subjected to immunoprecipitation with affinity-purified preimmune (pi) and immune (i) anti-Sro7 and anti-Myo2 antibody. The lane marked T represents the input of wild-type and myo2iq1,5 strains used.