Fig. 8.

DJ-1 knock-down did not reduce oxygen consumption rates (OCR) in astrocyte-enriched cultures. Mitochondrial respiration was analyzed by measuring OCR in living astrocytes using a Seahorse XF24 analyzer. The OCR values were then normalized to total astrocyte numbers, as assessed by GFAP in-cell Western analysis, on the same plates. Control (solid lines) and DJ-1 knock-down (dashed lines) astrocytes were compared at baseline, after treatments with 0, 20, or 40 nM rotenone (A, B), and then after the addition of 500 nM FCCP (A). In each graph, same-time/treatment comparisons were made between control and DJ-1 knock-down astrocytes by paired t-tests. Mean±SE shown, n=5. (A) There was no difference between the OCR of control and DJ-1 knock-down astrocytes under any treatment condition over a 1 h period. However, as expected, rotenone treatment reduced the OCR, FCCP stimulated the OCR, and rotenone reduced the FCCP-induced stimulation. The data in this graph are expressed as percent control relative to OCR/GFAP values from same-time no rotenone/no FCCP control wells on the same plates. (B) Extended assessments over 24 h did not show any significant differences between the OCR of control and DJ-1 knock-down astrocytes, but did show the expected rotenone-induced reduction in OCR. The data in this graph are expressed as percent control relative to OCR/GFAP values from same-time no rotenone control wells on the same plates. For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.