Fig. 3. Anti-oxidative effect of LMT497. (A, B) LMT497 reduced OGD/R-evoked oxidative stress in a cell culture system seen through DCF-DA assay. Cortical neurons were treated with LMT497 (10 µM) or Trolox (T; 10 µM) directly before the initiation of OGD. Cells were loaded with CM-H2DCF-DA for 10 minutes after reoxygenation, and intracellular ROS production was measured at 3 h after initiation of reoxygenation by an increase of fluorescence intensity (F.I). (A) Representative fluorescence images. Scale bar 50 µm. (B) Quantification of CM-DCF fluorescence. Horizontal bar, median; vertical box, interquartile ranges (Q1-Q3); and whiskers, minimum/maximum. n=6. ***p<0.001 (C-F) ROS scavenging activity of LMT497 in a cell free system. (C-E) The ORAC assay. (C, D) Representative time-dependent decay graphs of relative F.I in the presence of Trolox or LMT497 at different concentrations (-=untreated; •,○=6.25 µM; ▴,▵=12.5 µM; ✦,◊=25 µM; ▪,□=50 µM). (E) Best-fit lines between the net AUC (=AUCsample-AUCblank) and different concentrations of LMT497 of Trolox. n=4. (F) The DPPH assay. The dose-response curves of DPPH inhibition in the presence of LMT497 in comparison to vitamin C. n=4.
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