Fig. 9

In vitro data from mouse colonocytes suggests that NSAIDs, including phenylbutazone, induce oxidative stress and a subsequent cellular response known to occur when ER stress is induced by imbalanced redox homeostasis. A Fluorescent microscopic images of YAMC cells exposed to NSAIDs and H₂O₂, at the noted concentration, for 24 h prior to exposure to the ROS indicator CM-H2DCFDA. Ten images were taken from each treatment condition. B Average intensity of fluorescence for each treatment condition was significantly different that control cells (ANOVA) except the lowest concentration of H₂O₂. Graph represents data from 3 independent experiments. C Western blot of P62 protein from both the cytosolic (left blot) and nuclear (right blot) protein fractions of YAMC cells exposed to the NSAIDs at the indicated concentration for 24 h. Loading controls were the nuclear protein lamin A/C and the cytosolic protein GAPDH. D Graph of nuclear p62 represented as fold of DMF control from 3 independent experiments. DMF: dimethylformamide (0.04%), IB: ibuprofen (0.4 mM), PB: phenylbutazone (0.4 mM), IM: indomethacin (0.25 mM), H₂O₂ (0.5 or 0.1 mM as indicated)