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Fig. 1 | BMC Genomics

Fig. 1

From: A deficiency in SUMOylation activity disrupts multiple pathways leading to neural tube and heart defects in Xenopus embryos

Fig. 1

Depletion of SUMOylation activity in embryos injected with Gam1 mRNA. a Expression levels of Gam1 protein during embryogenesis. One-cell embryos were injected with mRNA (0.5 ng) encoding Gam1 with an N-terminal myc tag. Whole cell protein extract was prepared from embryos at the indicated Nieuwkoop–Faber stage and 25 μg taken for western blot analysis using anti-myc antibody. Lanes: 1, midblastula; 2, late blastula; 3, early gastrula; 4, early neurula; 5, late neurula; 6, water injected control. b In vitro SUMOylation assay. All assays contained Ubc9 (E2 enzyme), SUMO1, ATP, and substrate peptide (25 kDa) with (lane 1) or without (lane 2) purified E1 enzyme (500 nM) added. Samples were analyzed by western blot using an antibody specific for a 25 kDa SUMO substrate peptide. c Assays for E1 activity. One-cell embryos were injected with the indicated amount of Gam1 mRNA and allowed to develop to the indicated stage. Whole cell extract was prepared from 20 embryos and an equivalent amount of protein (25 μg) was used as a source of E1 enzyme for each assay

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