Fig. 3

POU6F2-AS1 facilitates de novo lipogenesis of CRC cell and PDO growth in vitro. (A) Heatmap of the differentially expressed genes after POU6F2-AS1 overexpression in CRC cells. (B) KEGG enrichment analysis showed the significantly enriched pathways after POU6F2-AS1 overexpression. (C) Gene Ontology (GO) analysis of these differentially expressed genes (DEGs) enriched in metabolic process showed the top 20 significantly enriched terms of biological process. (D, E, G) Representative confocal microscopy images of intracellular lipid droplets in CRC cells with POU6F2-AS1 knockdown or overexpression stained with BODIPY 493/503 dye (green). Nuclei were stained with DAPI (blue). (F, H) Representative images of intracellular lipid droplets in CRC cells with POU6F2-AS1 knockdown or overexpression stained with Oil Red O. (I) Heatmap showing liquid chromatography mass spectrometry (LC-MS) based FA analysis of HCT116 cells bearing control or POU6F2-AS1 knockdown. (J-N) qRT–PCR and western blotting analysis for mRNA and protein expression levels of key enzymes involved in FA synthesis (FASN, ACC1, and SCD1), FA β-oxidation (CPT1A) and FA uptake catabolism (CD36) in CRC cells with POU6F2-AS1 knockdown or overexpression. The data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001