MOG and ß-OHB generated ROS in insulin secreting cells.

<p>A. Both 200 µM MOG (squares) and 10 mM ß-OHB (triangles) increased ROS in INS-1 cells compared to basal (2 mM glucose) alone (circles). B. Area under curve of panel A. 200 µM MOG increased ROS generation in dissociated rat islets (C) compared to 3 mM glucose controls as measured with DCF fluorescence. D. 200 µM MOG (squares) and 10 mM ß-OHB (triangles) increased ROS generation in dissociated islet cells expressing the cytosolic HyPer protein compared to the basal (2 mM glucose) control as measured by changes in fluorescence ratio. E. Area under curve of D. A, B. n = 16 from 4 separate passages of cells. C. Results are typical of three separate experiments. D, E. Average of 14–33 cells from 3 separate animals. (A–C). ROS monitored for 1 hr in cells loaded with CM-H₂DCFA using a Tecan fluorescence platereader. (D–E). HyPer fluorescence ratio in dissociated islet cells monitored using a Zeiss 710 LIVE fluorescence microscope at 20× magnification. B and E, * p<0.005% compared to basal.