Fig. 4

PABPN1 regulates the 3’UTR lengths of ATP6AP2, TMEM97, CDCA2, CCND3, and AACS. A Abundance of transcripts with different 3’UTR lengths identified by 3’RACE assay. B Western blot assay measuring levels of PABPN1 targeted, Wnt signaling (ATP6AP2 and TMEM97), cell cycle (CDCA2 and CCND3), and lipid biosynthesis (AACS) related genes. GAPDH was used as a loading control. C Analysis of TOP/FOP luciferase reporter activity in cells stably transfected with ATP6AP2 or TMEM97 overexpressing vectors. D Flow cytometric analysis showing the cell cycle distribution of cells stably transfected with CDCA2 or CCND3 overexpressing vectors. E The neutral lipids content in cells stably transfected with AACS overexpressing vector. Neutral lipids were stained with BODIPY 493/503 dye (green). Nuclei were stained with Hoechst (blue). 200 × . F Quantification of phospholipid levels in AACS-overexpressing cells. G Quantification of triglyceride levels in AACS-overexpressing cells. H Proliferation of BC cells shown by colony formation assay. I Representative pictures of migrating and invading cells analyzed by Transwell assay. 200 × . Data are presented as the mean ± SD of three independent experiments. * P < 0.05