Fig. 7

887L interacts with 887S under normoxia and exhibits an inhibitory effect on 887S. A 887S was captured in the 887L-precipitated complex under normoxia, instead of hypoxia. 887S-P1 and 887S-P2: primer sets for 887S detection (n=2). WCL: Whole cell lysate. B 887S was captured in the 887L-precipitated complex under normoxia, instead of hypoxia. 887S-P1 and 887S-P2: primer sets for 887S detection (n=2). NF: Nuclear fraction. C DNMT1 ChIP assay showed the changes of DNMT1 recruitment to CA9’s CpG site in 887L knockdown cells under normoxia (n=3). D DNMT1 RIP assay showed the interaction of the DNMT1 and 887S in 887L knockdown cells under normoxia (n=3). E Representative image of RNA pull-down assay showed the association of DNMT1 and 887S in 887L knockdown cells under normoxia (n=3). IRE: a control RNA provided by Pierce RNA 3′ End Desthiobiotinylation kit. F Silencing of 887S RNA by antisense oligonucleotide (ASO) (ASO-887S) abolished the recruitment of DNMT1 to the CpG site of CA9 in 887L knockdown cells under normoxia, as determined by ChIP-qPCR assay (n=3). G Representative image of methylation status of the CpG sites on CA9 promoter in sh887L1 cells and control cells. PC: a primer set to detect the C to T conversion which is provided by the MSP kit. H A schematic illustration of the CA9 transcriptional regulation by interplay between 887S- and 887L-mediated pathways. Data are shown as means ± SEMs. P values are calculated using Student’s t test