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. 2018 May 8;9:615. doi: 10.3389/fpls.2018.00615

FIGURE 2.

FIGURE 2

SlPUB13 shows specificity toward group III E2s in in vitro ubiquitination assays. GST-SlPUB13 was tested against 34 purified tomato E2 proteins (Zhou et al., 2017) in in vitro ubiquitination assays to determine its specificity toward ubiquitin E2s. Tomato E2 protein used in each reaction is shown above the lane by their UBC number and the minus marker (–) denotes the absence of any E2 enzyme in the reaction. The presence of high molecular weight (MW) (>98 kD, size of GST-SlPUB13) smear of ubiquitinated proteins as detected by Western blot using anti-FLAG antibody indicates E2-E3 specificity. The numbers on the right denote the molecular mass of marker proteins in kilodaltons. This experiment was repeated two times with similar results.