Fig. 3.

PXL770 treatment improves VLCFA levels, restores mitochondrial function and increases compensatory gene expression in Abcd1 KO mouse glial cells. Mixed glial cells (astrocyte-enriched) were isolated from 1-day-old wt and Abcd1 KO mice. 10 days after culture cells were exposed to PXL770 at 5, 10, 25 or 50 μM. (A) Cells were exposed for 7 days to the treatment and VLCFA levels were analyzed by LC-MS after extraction of total lipids from pelleted cells. Data are mean ± S.E.M., n = 2 replicates for wild-type and n = 3 replicates for Abcd1 KO and PXL770. (B) Cells were exposed for 72 hours to the treatment and the OCR was measured using a Seahorse Analyzer. Bioenergetics parameters were evaluated by sequential additions of: oligomycin (1 μM), FCCP (0.5 μM) and Rotenone-Antimycin A (1 μM). Basal = first three measurements, ATP-linked = OCR drop after oligo addition, Maximal Oxidative capacity = after addition of FCCP. Data are mean ± SEM, n = 6 replicates. (C) Cells were exposed for 72 hours to the treatment and mRNA levels were evaluated by RT-qPCR, normalized by Rlp27 expression (no unit). Data are mean ± SEM, n = 3–5 replicates. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001 (Dunnett’s test versus untreated).