Research Paper Volume 15, Issue 18 pp 9572—9589

Figure 3. Silencing CF modifications with FUT8 siRNA rescues AECs from senescence and attenuates BLM-induced lung fibrosis. NT siRNA or FUT8 siRNA was used to transfect MLE₁₂ cells, which were subsequently treated or untreated with BLM for 48 hours. (A) To test the gene silencing effect, we use real-time polymerase chain reaction (RT-PCR) to show FUT8 mRNA expression in MLE₁₂ cells transfected with NT siRNA or FUT8 siRNA. ^**P < 0.01 compared to NT siRNA (Unpaired t-test). (B–G) Western blotting using anti-FUT8, anti-p21, anti-p16, anti-E-cadherin, anti-collagen III, and anti-β-actin antibodies in MLE₁₂ cells treated in different ways. Densitometric analyses of the bands were adjusted to be equal to β-actin. Data were presented as the mean ± SD. ^*P < 0.05. ^**P < 0.01. ^***P < 0.001. ^****P < 0.0001. (H, I) SA-β-gal staining reveals indicative pictures of different treated MLE₁₂ cells. Scale bar, 10 μm. Data were presented as the mean ± SD. ^*P < 0.05. ^**P < 0.01. (J–O) E-cadherin, p21^WAF1, p16^ink4a, collagen I, and collagen III expressions were measured via the immunofluorescence technique. Scale bar 25 μm. Data were presented as the mean ± SD. ^*P < 0.05. ^**P < 0.01. ^***P < 0.001. ^****P < 0.0001.