Fig 11.
ChCpc1 interacted with ChChk1 in Cochliobolus heterostrophus. (A) ChCpc1 interacted with ChChk1 in yeast two-hybrid assay. Serial dilutions of the yeast cells were plated on synthetic dropout (SD) medium that lacked histidine (H), leucine (L), and tryptophan (T) (SD- T/H/L). The interaction between pGADT7-T and ChChk1 was used as control. (B) Pull-down assay to verify the interaction between ChCpc1 and ChChk1. The fusion protein His-Cpc1 was detected in the eluted solution of GST-ChChk1 and His-ChCpc1 co-incubated mixture but not in that of glutathione S-transferase (GST) and His-ChCpc1. (C) Expression levels of ChARG1, ChARG4, ChATG5, ChATG7, ChATG8, and ChATG9 in WT and Δchk1 mutants under different culture conditions. CM: complete medium; MSX: L-methionine-DL-sulfoximine; MM: minimal medium; MM + MSX: minimal medium supplemented with 100 µM L-methionine-DL-sulfoximine (MSX). GraphPad Prism program’s t-tests and multiple t-tests were used for the analysis of significant differences. The bars indicate the standard error of means of three replications.