Research Paper Volume 11, Issue 5 pp 1356—1388

Alternative polyadenylation dependent function of splicing factor SRSF3 contributes to cellular senescence

Figure 5. SRSF3-KD induced 3′ UTR shortening of PTEN produces more protein and contributes to senescence-associated phenotypes. (A) Dual luciferase assay for shorter (S) and longer (L) 3′ UTRs of PTEN in 293T (left) and HUVEC (right) cells. (B) RNA degradation curve of actD-inhibited transcription cells detected by qRT-PCR using primers specific for longer 3′ UTR (L) or targeting common region shared by longer and shorter 3′ UTR (T) of PTEN. (C, G) qRT-PCR indicated overexpression of PTEN (PTEN_OE and Pten_OE) led to decreased expression of cell proliferation marker MKI67 in 293T (C) and MEF (G) cells. (D, H) SA-β-Gal staining for 293T (D) and MEF (H) cells when overexpressing PTEN. (E, F, I, J) CCK-8 assay (E, I) and cell cycle analysis (F, J) for cells with or without overexpression (OE) of PTEN in 293T (E, F) and MEF (I, J) cells. *** and * mean P value less than 0.001 and 0.05, respectively (t-test).