Fig. 3.

ZTL, GI, and TOC1 protein levels are regulated by UBP12 and UBP13. a, c, e The protein levels of HA-tagged GI driven by native promoter (pGI::GI-HA), ZTL and YFP-tagged TOC1 driven by the TOC1 promoter (TOC1 minigene or TMG) in the wild type (Col-0), ubp12-1, or ubp13-1 mutants under diurnal conditions (12 h light/12 h dark) were detected by immunoblotting. The samples from 0 h to 12 h after dawn were harvested in light, and the samples from 16 h and 20 h after dawn were harvested in the dark (indicated by gray shading). The relative protein levels were quantified by normalization to actin. The Col-0 or ztl-4 samples were used as negative controls for the antibodies. Plots represent the average protein levels from three biological replicates, and the error bars represent standard deviation. Compared to wild type, the levels of ZTL proteins in the ubp12-1 and ubp13-1 were below the linear range for quantification. In the ztl-4 sample, the anti-ZTL antibody recognizes a non-specific band close to the size of endogenous ZTL in the long-exposure blots. b, d, f The relative mRNA levels of GI-HA, ZTL, or TOC1-YFP from the same time course samples were measured by qRT-PCR. The source data are provided as a Source Data file. Blot images were cropped from their original size, which can be found in Source Data file