SAHA treatment upregulates Abcd2 and Abcd3 levels in Abcd1-deficient U87 astrocytes and B12 oligodendrocytes.

There was no change in Abcd1 protein levels (A-i and E-i) and mRNA expression (A-ii and E-ii) in Abcd1-deficient U87 astrocytes and B12 oligodendrocytes treated with SAHA. Abcd2 (B and F) and Abcd3 (C and G) protein levels (i) and mRNA expression (ii) in control, NT and Abcd1-deficient U87 astrocytes and B12 oligodendrocytes respectively (n = 3) treated with SAHA dose-dependently. Protein levels of the peroxisomal transporters Abcd1 (A-i and E-i), Abcd2 (B-i and F-i) and Abcd3 (C-i and G-i) were analyzed by Western analysis of peroxisomal membrane fractions obtained by carbonate treatment (membrane preparation containing integral membrane proteins), as indicated in Methods section. Na⁺/K⁺-ATPase (plasma membrane protein) was used as indicator of protein loading for plasma membrane fractions. Abcd1 (A-ii and E-ii), Abcd2 (B-ii and F-ii) and Abcd3 (C-ii and G-ii) mRNA levels were determined by qRT–PCR and normalized to GAPDH. Data are represented as mean±SD. ^@P<0.001 Abcd1-Lenti compared to control or NT cells; **P<0.001 SAHA (5.0 µM) treatment compared to Abcd1-Lenti; *P<0.005 SAHA treatment compared to Abcd1-Lenti; ^@@P<0.001 SAHA treatment compared to Abcd1-Lenti; NS, non-significant; SA, SAHA.