Fig. 5.

LINC00930 functions as a link between RBBP5/GCN5 and PFKFB3. a Western blot assay was performed to measure PFKFB3 level after transfection of LINC00930-overexpressing plasmid, RBBP5 or GCN5 siRNAs in CNE1 cells. b Luciferase reporter vector was generated by inserting the promoter region (− 2000 bp to + 200 bp) of the PFKFB3 gene. The reporter vectors were then cotransfected into CNE2 cells with LINC00930 shRNA, RBBP5 or GCN5 siRNAs. Cells were harvested for luciferase activity assay. c ChIP-qPCR was performed to evaluate the enrichment of H3K4me3 or H3K9ac in different promoter regions of PFKFB3 in CNE2 cells after knocking down LINC00930. d Real-time PCR of the ChIP samples was applied to detect the binding efficiency of RBBP5 or GCN5 to the PFKFB3 gene promoter (− 800 bp to − 600 bp) after transfection of RBBP5, GCN5, or LINC00930 siRNAs in CNE2 cells, respectively. e Real-time PCR of the ChIP samples was utilized to measure the H3K4 trimethylation or H3K9 acetylation and H3K27 trimethylation levels of the PFKFB3 gene promoter (− 800 bp to − 600 bp) after knockdown of RBBP5, GCN5, or LINC00930 in CNE2 cells, respectively. f ChIRP assays of the enrichment of LINC00930 and PFKFB3 promoter (− 800 bp to − 600 bp) in both even and odd probes pools relative to control LacZ probes set in 5-8F and CNE2 cells. Data in b, c, d, e & f are presented as mean ± SEM of three independent experiments. The p-value was determined by a two-tailed unpaired Student’s t test. * p < 0.05; ** p < 0.01