Research Paper Volume 15, Issue 1 pp 70—91

Figure 3. Inhibition of Smyd2 alleviates Ang II-induced senescence associated phenotypes in vitro. (A, B) SA-β-gal staining (A) and EdU incorporation (B) assays in Ang II-induced RAECs with LLY-507 (3 μM) pretreatment. Representative staining images are shown in the figure left, and the statistical analysis of positive cells is shown in the figure right. Scale bars, 100 μm. (C) Expressions of senescence markers (p53, p21 and p16), pro-inflammatory molecules (VCAM-1 and COX-2) and DNA damage markers (p-Chk2 and γH2AX) were detected by western blot analysis upon LLY-507 pretreatment in Ang II-induced RAECs. Representative western blot images are shown on the left, and the statistical analysis of relative protein expressions is shown on the right. GAPDH was used as the loading control. (D) The mRNA levels of Nos2 (encoding the iNOS protein) in Ang II-induced RAECs with LLY-507 pretreatment was examined by RT-qPCR. (E, F) Immunofluorescence double staining of Smyd2 and the senescence markers (p53 and p16) upon LLY-507 pretreatment in Ang II-induced RAECs. Scale bars, 100 μm or 50 μm. Data are presented as the mean ± SEMs, ^*p < 0.05, ^**p < 0.01, ^***p < 0.001, each acquired from three individual experiments (n = 3).