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. 2018 Apr 18;9:769. doi: 10.3389/fmicb.2018.00769

FIGURE 8.

FIGURE 8

MoUbp14 is responsible for deubiquitination of MoFbp1 and MoPck1. (A) MoUBP14 deletion mutant increased in total protein ubiquitination level. Total lysates of P131 and the ΔMoubp14 mutant were subjected to SDS-PAGE and immunoblotted using anti-ubiquitin antibodies to detect total Ub levels. (B) Ubiquitination levels of MoFbp1 and MoPck1 are significantly increased in the ΔMoubp14 mutant. MoFBP1:Flag and MoPCK1:Flag fusion constructs were transformed into P131 and the ΔMoubp14 mutant, respectively. Corresponding transformants were analyzed by western blotting with anti-Flag antibody. Staining of total protein with Ponceau S confirmed equal loading. (C) MoUbp14 is required for MoFbp1 and MoPck1 degradation in short time starved condition. 12 h-starved wild type and the ΔMoubp14 mutant strains, respectively, expressing MoFBP1:Flag (or MoPCK1:Flag) were shifted to glucose for 2 h. Total extracts were subjected to SDS-PAGE and immunoblotted using anti-Flag antibodies. Staining of total protein with Ponceau S confirmed equal loading.