Figure 4.

Mitochondrial bioenergetics in adenosine monophosphate activated protein kinase- (AMPKα1-) deleted Abcd1-knockout (KO) primary mixed glial cells. (a) Proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) protein and mRNA levels were significantly reduced in Abcd1-KO cells when deleted for AMPKα1. (b) Abcd1-KO mice mixed glial cells (1.5 × 10⁴ cells/well) were plated in XF⁹⁶V3-PS cell culture plates (Seahorse Bioscience) and deleted for AMPKα1 as described in Section 2. Mitochondrial oxygen consumption rate (OCR) was measured by sequential addition of oligomycin, FCCP, and antimycin/rotenone to measure basal and FCCP-uncoupled OCR. (c) Basal and FCCP-linked OCR was similar between wild type (WT) and Abcd1-KO mixed glial cells but decreased in Abcd1-KO mixed glial cells deleted for AMPKα1. (d) A parallel 96-well plate was used for adenosine triphosphate (ATP) measurement. Results represent the mean ± SD of triplicates from three different experiments. ^*** P < 0.001. NS: nonsignificant.