Figure 6.

hSav1 changes the levels of phosphorylation of specific substrates by Mst2. A, plasmids encoding a HA-Mst2 or FLAG-Mob1 were transiently co-transfected with or without HA-hSav1 in HEK293T cells. Normalized cell lysates were analyzed by Western blotting for protein expression with antibodies that recognize each epitope tag (HA or Flag), the two phosphorylation sites of Mob1 (pT12 or pT35), or β-Actin as a loading control. Representative blots from one experiment (top) and a scatter plot of three replicates (bottom) are shown. Data are plotted as pThr-12 or pThr-35 Mob1 intensity divided by total FLAG-Mob1 intensity (p values for both > 0.05). B, the same experiment as in panel A but with a plasmid encoding HA-Lats1 instead of FLAG-Mob1 and antibodies specific to the hydrophobic motif phosphorylation of Lats1 (pT1079) instead of those specific to phosphorylated Mob1. Representative blots from one experiment (top) and a scatter plot of seven replicates (bottom) are shown. Data are plotted as pThr-1079 Lats1 intensity divided by total Myc-Lats1 intensity (p values ≤ 0.001).