Fig. 6
miR-500a-5p expression is up-regulated via the YY1/p300/HDAC2 complex. a In vivo co-immunoprecipitation assays showing the presence of a complex containing YY1, HDAC2 and p300. HA-tagged p300 plasmid was transfected into CRC cells. Immunoprecipitation was performed with anti-HA (p300) antibody, and pre-immune normal mouse immunoglobulin G (nm IgG) was used as a control. Western blot analysis was performed with an anti-HDAC2 (α-HDAC2) and a YY1 antibody. b HDAC2 plasmid was transfected into CRC cells. Cell lysates were immunoprecipitated using an anti-HDAC2 antibody or the control antibody normal mouse immunoglobulin G (nm IgG). Western blotting was performed with anti-HA (p300) and YY1 antibodies. c and d Whole cell lysates were immunoprecipitated using an anti-p300 (c) or anti-HDAC2 (d) antibody or the control antibody normal nm IgG. Western blotting was performed with HDAC2 or p300 and YY1 antibodies. e Gene expression studies by qRT-PCR in SW620 and LoVo cells. Student’s t test; **P < 0.05, ***P < 0.01 and ****P < 0.001 between vector and gene. f The miR-500a-5p promoter construct was co-transfected with vector, YY1, HDAC2, and p300 either alone or in combination with HDAC2 + p300, YY1 + p300, and HDAC2 + p300 + YY1 in CRC cells. Luciferase assays were performed. Values are means ± SD (n = 3). Student’s t test; ***P < 0.01 and ****P < 0.001 between vector and gene. g and h Binding of the transcription factor YY1 to miR-500a-5p was analysed with anti-YY1 ChIP using qPCR. Student’s t test; **P < 0.05 and ***P < 0.01 between vector and gene. All figures are representative of three independent experiments
