Figure 4

ROS and TRX activity in response to glucose and p38 MAPK inhibition. A) TXNIP RNA message levels were detected by semi-quantitative PCR in MDA-MB-231 cells grown either in 5 or 20 mM glucose chronically prior to plating. Average relative levels as compared to control β-actin RNA of TXNIP and TRX RNA messages from triplicates are represented. For inhibition of the p38 MAP kinase cells grown at 20 mM were pre-treated for 24 h with 20 μM SB203580. B) MDA-MB-231 cells were grown either in 5 or 20 mM glucose chronically prior to plating. Cells were assessed for ROS levels by DCFDA fluorescence staining and flow cytometry as shown in the inlet. For inhibition of the p38 MAP kinase cells grown at 20 mM glucose were treated for 24 h with 20 μM SB203580. Average mean fluorescence from triplicates is expressed per each group of cells. B) MDA-MB-231 cells were grown either in 5 or 20 mM glucose chronically prior to plating. Cells were assessed for TRX activity by the insulin disulfide reducing assay as described and the average OD 410 readings from triplicates are shown for each group of cells. For inhibition of the p38 MAP kinase cells grown at 20 mM were pre-treated for 24 h with 20 μM SB203580.