Research Paper Volume 12, Issue 5 pp 4322—4336

Sirt1 antisense long non-coding RNA attenuates pulmonary fibrosis through sirt1-mediated epithelial-mesenchymal transition

Figure 5. Knockdown of lncRNA sirt1 antisense (AS) reverses the protective effects of astragaloside IV (ASV) on TGF-β1-induced fibrogenesis. (A) Sirt1 AS expression in RLE-6TN cells treated with various concentration of ASV for 48 h, as indicated. * P<0.05. (B) Sirt1 AS expression in TGF-β1 treated RLE-6TN cells upon ASV treatment. # P<0.05 vs. control; ## P<0.05 vs. TGF-β1. (C) CCK-8 assay was used to detect the cell viability of RLE-6TN cells treated with 10 ng/ml TGF-β1, TGF-β1+100μM ASV, TGF-β1+ASV+sh-Scramb, TGF-β1+ASV+sh-sirt1 AS. (D) Western blotting analysis and quantitative analyses of EMT related markers α-SMA, E-cadherin, collagen1 and fibronectin1 in RLE-6TN cells treated with TGF-β1, TGF-β1+ASV, TGF-β1+ASV+sh-Scramb, TGF-β1+ASV+sh-sirt1 AS. (E) Immunofluorescence staining showing the overlap of α-SMA and E-cadherin in RLE-6TN cells with indicated treatment. (F) Transwell assay was performed to investigate the migration ability of RLE-6TN cells with indicated treatment. # P<0.05 vs. control group; ## P<0.05 vs. TGF-β1; $ P<0.05 vs. TGF-β1+ASV+sh-Scramb.