Fig. 3

MEN1 knockdown leads to increased JunD binding and loop structure formation in the MYC locus in AR-independent PCa cells. a ChIP-qPCR analysis using anti-menin to evaluate the binding of menin to the MYC 5′ enhancer (− 67 kb) and promoter in DU145 and PC3 cells treated with siCtrl or siMEN1(1) + (3). Chr1 served as a negative control. b ChIP-qPCR analysis using anti-JunD to detect the binding of JunD to the MYC 5′ enhancer and promoter in DU145 and PC3 cells treated with siCtrl or siJunD. c ChIP-qPCR analysis to assess the effect of MEN1-KD on the level of JunD recruitment to the MYC 5′ enhancer in DU145 and PC3 cells. d Primer locations for qPCR of 3C and ChIP-3C analyses. e 3C detection results (DP1-DP2 fragment) and control (CP1-CP2) by PCR in siCtrl or siMEN1(1) + (3)-transfected DU145 cells upon ligation or non-ligation. ChIP-3C qPCR analysis detecting menin ChIP-qPCR analysis showing menin binding (f) and JunD binding (g) to the MYC 5′ enhancer, MYC promoter and the looping fragment (F3-F6) in DU145 cells transfected with siCtrl or siMEN1(1) + (3) under ligation (right panel) or non-ligation (left panel) conditions, P1-P2 and P7-P8 were used as negative controls. Representative blots of three independent experiments