Fig. 5

Phosphorylation of LSD1 by PLK1 promotes its chromatin release during mitosis. a Nocodazole treatment induced LSD1 release from chromatin. HeLa cells were asynchronized or synchronized at metaphase by single thymidine block followed by nocodazole treatment for 12 h and then subjected to chromatin fractionation. Soluble cytosol and nuclear soluble fraction, chromatin chromatin-enriched fraction. Relative ration of the band intensity of total LSD1/GAPDH in the soluble fraction or total LSD1/H3 in the chromatin-enriched fraction was presented in (b) and (c) as well. b PLK1 inhibitor treatment blocked nocodazole-induced chromatin release of LSD1. HeLa cells were prepared as in a except that a fraction of thymidine-nocodazole synchronized cells were treated with BI2536, a PLK1 inhibitor, 2 h before harvest. c Phosphorylation-defective mutant LSD1 (S126A) failed to release from chromatin during mitosis. HEK293 cells stably expressing HA-LSD1 or HA-LSD1(S126A) were subjected to chromatin fractionation