Fig. 4

MoCsn5 regulates the ubiquitin‒proteasome pathway in rice blast fungus. A Ubiquitination levels of 70-15, ΔMocsn5 and Mocsn5-C at 0 h and 3 h after MG132 induction. Hyphae were grown in liquid CM for 36 h and then shifted to CM supplemented with 20 μM MG132 (a proteasome inhibitor, a tool for studying cellular degradation of the ubiquitin‒proteasome pathway) for 0 h and 3 h. B Transcript levels of MoSKP1 (MGG_04978), MoCULLIN1 (MGG_07145), MoCULLIN3 (MGG_07731) and MoCULLIN4b (MGG_14763) in 70-15 and ΔMocsn5. The error bars represent the standard deviations. Tukey’s test was used to determine significance. **P < 0.01. C The interaction between MoCsn5 and MoCullin3 was detected by yeast two-hybrid assays. pGADT7-T and pGBKT7-53 were used as positive controls. pGADT7-T and MoCsn5-BD and pGBKT7 and MoCullin3-AD served as two pairs of negative controls. D The relationship between MoCsn5 and MoCullin3 in vivo was examined by coimmunoprecipitation assays. The MoCullin3-Flag bands were detected following MoCsn5-GFP immunoprecipitation