Figure 7.
Ypk1/2 is required for normal hyperphosphorylation of Net1 in vivo. (A)NET1∆328-3xHA and NET1∆328-3xHA ypk2∆ ypk1-as1 cells grown overnight in YPD were released from a G1 phase arrest at 25°C into YPD medium containing benomyl. 3MOB-PP1 was added to both cultures to a final concentration of 10 uM at 40 min after release. Samples were taken at the indicated time intervals to assay for Net1∆328-3xHA and Clb2 by western blot. An anti-Nap1 antibody was used as a loading control. (B and C) Samples from multiple biological replicates of the experiment shown in A were analyzed to determine the percentage of cells undergoing bud emergence (B) and to measure median cell size with a Coulter Counter (C). Error bars indicate SEM for three biological replicates. Source data are available for this figure: SourceData F7.

Ypk1/2 is required for normal hyperphosphorylation of Net1 in vivo. (A) NET1 ∆328 -3xHA and NET1∆328-3xHA ypk2∆ ypk1-as1 cells grown overnight in YPD were released from a G1 phase arrest at 25°C into YPD medium containing benomyl. 3MOB-PP1 was added to both cultures to a final concentration of 10 uM at 40 min after release. Samples were taken at the indicated time intervals to assay for Net1∆328-3xHA and Clb2 by western blot. An anti-Nap1 antibody was used as a loading control. (B and C) Samples from multiple biological replicates of the experiment shown in A were analyzed to determine the percentage of cells undergoing bud emergence (B) and to measure median cell size with a Coulter Counter (C). Error bars indicate SEM for three biological replicates. Source data are available for this figure: SourceData F7.

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