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. 2014 Jul 16;34(29):9574–9589. doi: 10.1523/JNEUROSCI.4302-13.2014

Figure 1.

Figure 1.

CREB is sumoylated by PIAS1 in vitro and in HEK293T cells. A, Alignment and comparison of CREB1α and CREB1Δ isoforms showing that CREB1Δ is 14 aa shorter than that of CREB1α (aa 88–101), and that Ser119 of CREB1Δ corresponds to Ser133 of CREB1α. B, In vitro SUMOylation assay and Western blot showing PIAS1 SUMOylation of CREB. “a” and “b” indicate the sumoylated form of the CREB bands. C, Co-IP experiment showing the association between PIAS1 and CREB in HEK293T cells with or without the cotransfection of Myc-SUMO1 or Myc-SUMO1 (ΔGG). The quantified results are shown on the bottom. D, Transfection of different plasmids to HEK293T cells and Western blot showing PIAS1 SUMOylation of CREB in HEK293T cells. E, Confirmation of PIAS1 SUMOylation of CREB in HEK293T cells by IP and Western blot against CREB and SUMO1. endo-CREB, Endogenous CREB; ΔGG, C-terminal diglycine motif truncated SUMO1. Data are expressed as mean ± SEM. **p < 0.05; **p < 0.01; #p < 0.001. Experiments were performed in triplicate.