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. 2023 Oct 26;18(10):e0293311. doi: 10.1371/journal.pone.0293311

Fig 3. Pharmacological inhibition of ROS and phospho-Erk(1/2) ameliorates Bu-induced endothelial cell damage.

Fig 3

SK-HEP1 cells were pretreated with N-acetyl cysteine (NAC) (1mM) or rosiglitazone (1mM) or metformin (2mM) or U0126 (10uM). One hour post-treatment, the cells were incubated with Bu (300uM). Eight hours post-treatment, the cells were analyzed for ROS levels by measuring DCFDA fluorescence by flow cytometry. A. representative histogram overlay of inhibition of ROS assay. B. Graph showing MFI levels indicating ROS levels post-drug pretreatment. C and D. Western blot images showing levels of cleaved PARP, cleaved caspase 3, phospho-Erk(1/2) post inhibitors pretreatment. E, F, and G. Graphs showing relative levels of the proteins normalized to GAPDH by densitometric analysis using ImageJ software. H. Apoptosis percentages of SK-HEP1 cells pretreated with metformin (2mM) for 24 hours and Bu for 48h. Data shown are mean± SD of three independent experiments in triplicates. The P-value was calculated by the Mann-Whitney U test. * p<0.05 vs. mock-treated cells.