Fig. 2

Effects of 5-methoxyflavone on the LPS-mediated NOX4/TLR4/NF-κB/P38 MAPK pathway in BEAS-2B cells. (A, B) BEAS-2B cells were stimulated with LPS (10 μg/mL) in the presence or absence of 5-methoxyflavone for 12, 24 and 48 h. Then, cells were incubated with 5 μM DCFH-DA at 37°C for 30 min. The levels of ROS in LPS-stimulated BEAS-2B cells treated with 5-methoxyflavone were detected by fluorescence microscopy (A) and flow cytometry (B). (C) BEAS-2B cells were stimulated with LPS (10 μg/mL) for 2, 6, 12, 24 and 48 h. Expression of NOX4, TLR4, P-P65 and P-P38 was detected by western blotting. (D, E) Expression of NOX4 (D) and TLR4 (E) in LPS-stimulated BEAS-2B cells treated with 5-methoxyflavone was detected by western blotting. (F) Expression of P-P65 and P-P38 in LPS-stimulated BEAS-2B cells treated with 5-methoxyflavone were detected by western blotting. The band intensity of targeted proteins was determined by ImageJ software and was normalized to the intensity of GAPDH. ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 versus the control group; ^*p < 0.05, ^**p < 0.01 ^***p < 0.001 versus the LPS group