Research Paper Volume 16, Issue 9 pp 7752—7773

Figure 7. HOTAIR affects CRC proliferation through uridine bypass via EZH2/UPP1 axis. (A) A hierarchically clustered heatmap of differentially expressed genes in CRC cells after transfection of si-HOTAIR or NC-siRNAs. (B) Ten representative genes expression levels in CRC cells depleted of HOTAIR. (C) The knockdown and over-expressing efficiency of UPP1 were detected by RT-qPCR. (D) RIP assays were performed in CRC cells to show HOTAIR co-immunoprecipitation with EZH2, LSD1, Ago2 and SUZ12. (E, F) EZH2 and UPP1 mRNA and proteins level in CRC cells transfected with si-EZH2 or siRNA-NC by qRT-PCR and western blot analysis. (G) EZH2 occupancy on the UPP1 promoters was upregulated by HOTAIR knockdown by ChIP-qPCR assay. (H) UPP1 was highly expressed in CRC tissues compared with normal tissues through TCGA. (I) Reaction catalysed by UPP1 proteins. (J) Cell growth assays of HCT116 control cells in glucose-free media in the presence of 10 mM of either glucose, uridine or ribose. (K) Colony-forming assays assessed CRC cell proliferation in the presence of si-HOTAIR, sh-UPP1, OE-UPP1, glucose, uridine or ribose.