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. 2017 Dec 4;216(12):4041–4052. doi: 10.1083/jcb.201703096

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© 2017 Ribet et al.

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Figure 2. — SUMOylation of human septins. (a) HeLa cells were cotransfected with WT His₆-SUMO1, 2, or nonconjugatable (ΔGG) mutants and HA-tagged septins. Cell lysates were then subjected to denaturing His pull-down, and the presence of SUMOylated septins was assayed by immunoblot analysis using anti-HA antibodies (asterisks represent nonspecific binding of un-SUMOylated septins to nickel–nitrilotriacetic acid beads). Input fractions are shown as controls. (b) Immunoblot analysis, using anti-SEPT7 antibodies, of His pull-down proteins from HeLa cells transfected with WT or nonconjugatable (ΔGG) His₆-SUMO1. Input fractions are shown as control. (c) Immunoblot analysis of His pull-down proteins from synchronized HeLa cells transfected with WT His₆-SUMO1. Anti-RanGAP1 antibodies were used to monitor pull-down of SUMOylated proteins. Antiphosphorylated histone H3 antibodies (phos-H3) were used to control cell cycle synchronization.