Fig 2. Homologous scaffolds Atg11 and Atg17 compete for binding to Atg9.
(A) Schematic drawing illustrates the addressed question. Predicted CC regions of Atg11 and Atg17, and interaction sites with other Atg proteins are indicated. Sequence alignment by ClustalW suggests that Atg17 is homologous to Atg11122-536. (B) IP of myc-tagged Atg11 and co-IP of HA-tagged Atg9 from lysates of WT (−), atg13Δ, atg17Δ, atg29Δ, and atg31Δ cells (log phase). Lysate of nontagged atg11 served as control. Atg11 and Atg9 were detected by α-myc and α-HA immunoblots, respectively. The chart shows corresponding quantifications of Atg9 intensities normalized to the WT and corrected for the total amount of Atg11. (C) IP of myc-tagged Atg11 and co-IP of HA-tagged Atg9 as in (B) but from lysates of starved cells. (B and C) Data are presented as mean values ± SD of n = 3 independent experiments. P values were calculated using a two-tailed Student t test (*P < 0.05, **P < 0.01, ***P < 0.001). Uncropped original blots are shown in S5 Fig. CC, coiled–coil; HA, hemagglutinin; IP, immunoprecipitation; WT, wild-type.