Research Article

Poldip2/Nox4 Mediates Lipopolysaccharide-Induced Oxidative Stress and Inflammation in Human Lung Epithelial Cells

Figure 5

Downregulation of Poldip2 decreased LPS-induced oxidative stress and inflammation. (a, b) Confluent A549 and Beas-2B cells were transfected with the lentivirus downregulation vector for 72 h, and the efficiency of Poldip2 downregulation was verified by Western blotting and quantitatively analyzed. Following transfection, they were treated with LPS (10 μg/mL for A549 cells and 1 μg/mL for Beas-2B cells) for 12 h. (c, d) The protein levels of Nox4, HO-1, and COX-2 were detected by Western blotting and quantitatively analyzed. (e–g, i) The expression of Nox4 in the transfected cells was detected by immunofluorescence analysis and laser confocal microscope. The cells were stained with Nox4 (green) and counter stained with DAPI (blue) (magnification of 400x). (h, j) The NADPH enzymatic activity in cells was detected by a spectrophotometer. (k, l) The production of intracellular ROS was detected by flow cytometry. (m–r) The expression levels of PGE2, TNF-α, and IL-1β in the supernatant were detected by ELISA. Bars are of 3 independent experiments. vs. shCont, vs. shCont, vs. shCont, vs. , vs. , and vs. .
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