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Fig. 8. Suppression of Pma1-10 ubiquitylation independent of slow ER export. Cells bearing HA-tagged Pma1 were induced for 2 hours at 37°C and then chased for another 2 hours. (A) Indirect immunofluorescence localization of HA-Pma1-10 in yvh1 cells. Cells were fixed and stained with anti-HA and anti-Kar2, as described in Materials and Methods. Digital images were captured at the same exposure time and adjusted with Adobe Photoshop at the same settings. (B) Pma1-10 ubiquitylation, but not Pma1-7 ubiquitylation, is prevented in yvh1. Wild-type (L3852) and yvh1 (KKX15-1B) bearing pS3 (MET25-HA-pma1-7) or pKK5 (MET25-HA-pma1-10) were induced for 2 hours at 37°C. Anti-HA immunoprecipitates were analyzed by western blotting with anti-ubiquitin and anti-HA antibodies. Arrow indicates position of non-ubiquitylated Pma1.





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