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. 2010 Apr 13;29(11):1803–1816. doi: 10.1038/emboj.2010.63

Figure 4.

Figure 4

AO domain of LSD1 is responsible for its interaction with Snail1. (A) The cartoon and schematic diagram shows the structure of LSD1 and the different deletion constructs used in this study. (B, C) HA-tagged Snail1 and Flag-tagged full-length or deletion mutants of LSD1 were co-expressed in HEK293 cells treated with MG132 for 6 h. After immunoprecipitation, bound Snail1 (B) and LSD1 (C) were examined by western blotting. (D) GST-tagged full-length or deletion mutants of LSD1 were incubated with lysate from HEK293 cells expressing Snail1. The GST pull-down complex was eluted with SDS–PAGE buffer and about one-tenth of these elutents were analysed for the association of Snail1 by western blotting (bottom panel). The rest of the elutents were examined for the presence of purified GST–LSD1 by Coomassie staining (top panel). (E) Flag-tagged AO domain, AO domain without Tower (AOΔTower) or Tower domain of LSD1 was co-expressed with Snail1 in HEK293 cells treated with MG132 for 6 h. After immunoprecipitation, the bound deletion mutants of LSD1 and Snail1 were examined by western blotting.