Fig. 6

SATB2-AS1 regulates DNA demethylation and H3K4me3 enrichment of the SATB2 promoter by recruiting WDR5 and GADD45A. a, SATB2-AS1 targeted probes and negative Laz targeted probes were used for ChIRP assay. Purified DNA and RNA was analyzed by qRT-PCR respectively. ‘Odd’, ‘Even’ and ‘LacZ’ denote the odd- and even-ranked corresponding probes against SATB2-AS1 and probes against bacterial LacZ, respectively. Primer locations of C4, C7 and C9 are shown at the bottom of the Fig. 5f. b, ChIP-qPCR assays were performed to detect H3K4me3 occupancy at the SATB2-AS1 promoter region with indicated treatment. Primer locations are shown at the bottom of the Fig. 5f. c, Bisulfite sequencing was performed to evaluate the changes of CpG island methylation statuses in R3 and R4 regions in CRC cells after SATB2-AS1 knockdown. d, SATB2 levels were determined by qRT-PCR in CRC cells with indicated treatment. e, SATB2, E-cadherin and Vimentin expression were detected by western blot in CRC cells with indicated treatment. f, Schematic of the proposed mechanism of SATB2-AS1 in CRC. *, P < 0.05; **, P < 0.01 and ***, P < 0.001