Fig. 2: CXXC5 and TET2 occupancy at ARBS-gained CpGi-positive sites in ENZ-resistant CRPC cells.

a Heatmap showing RNA-seq read intensity of 12 CXXC domain genes in C4-2^CON and C4-2^ENZ-R cells. b UCSC tracks showing RNA-seq signal profiles of CXXC4 and CXXC5 expression in C4-2^CON and C4-2^ENZ-R cells. c Western blotting showing the AR, CXXC5, CXXC4, and TET2 protein level in cells expressing control or AR-specific shRNAs. ERK2 was used as a loading control. d In vitro pulldown analysis of CXXC5 interaction with AR and TET2 using proteins purified from baculovirus-insect Sf9 cell expression system. e Co-IP of endogenous proteins shows AR interaction with CXXC5, TET1, TET2, and TET3 in C4-2^CON and C4-2^ENZ-R cells. f, g Heatmap (integration of all replicates) shows ChIP-seq read intensity of CXXC5 and TET2 at ARBS-gained regions in C4-2^CON and C4-2^ENZ-R cells. h, i CXXC5 or TET2 was knocked down by specific shRNAs in C4-2^CON and C4-2^ENZ-R cells and the cell proliferation was measured by SRB assay. Data are represented as means ± s.d. (n = 6 replicates/group). Statistical significance was determined by two-way ANOVA. Experiments in c–e were repeated twice.