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. 2020 Jul 28;10:1234. doi: 10.3389/fonc.2020.01234

Figure 4.

Figure 4

LSD1 regulates LEF1 by forming a complex with β-catenin. (A) The top 15 pathways of RNA-Seq result. (B) Venn diagram showed that the overlapping gene among the top five pathways was LEF1. (C) Confocal microscopy was used to determine the LEF1 protein expression in T24 cells with or without LSD1 knockdown. (D) Expression of LEF1 analyzed by RT-qPCR after transfection with shRNA or siRNAs for LSD1 in T24 and BIU87 cells. Expression of LEF1 analyzed by Western blotting after transfection with shRNA or siRNAs for LSD1 in T24 and BIU87 cells. (E) EMT markers were determined by Western blotting in T24 and BIU87 cells with LSD1 knockdown when the expression of LEF1 was rescued by transient transfection of a plasmid expressing LEF1. The data represent means ± S.D. *P < 0.05. (F) T24 and BIU87 cells were lysed and subjected to co-immunoprecipitation with LSD1 and β-catenin antibodies. (G) 293T cells were transfected with indicated plasmids. co-immunoprecipitation was conducted by using Flag and HA antibodies, immunoblot was analyzed with the above antibodies. (H) T24 cells with or without LSD1 knockdown were subjected to CHIP assays with LSD1 antibody. (I) T24 cells with or without β-catenin knockdown were subjected to CHIP assays with LSD1 antibody. (J) T24 cells with or without LSD1 knockdown were subjected to CHIP assays with H3K4me1, H3K4me2, H3K9me1, and H3K9me2 antibodies.