C0979

4.1G (1:100; kindly provided by E. Peles) AnkG (1:100; kindly provided by M. Rasband), APP (1:1000; MAB348, Merck Millipore, Massachusetts, USA), ATG5 (1:100; pab50264 Covalab, France), CASPR (mk, m; 1:1000; clone K65/35 NeuroMab, California, USA), CASPR2 (1:500; kindly provided by E. Peles), Catalase (1:200; C0979 Sigma-Aldrich, Missouri, USA), CD3 (1:150; MCA1477 Serotec, MorphoSys, Germany), EEA1 (1:300; ab2900 Abcam, Cambridge, United Kingdom), K_v1.1 (1:50; sc11184 Santa Cruz Biotechnology, Texas, USA), K_v1.1 (1:50; clone 20/78 NeuroMab), K_v7.2 (1:2000; PA1-929 ThermoSicentific), LAMP1 (WB, 1:400; IF, 1:200; IEM, 1:200; 553792 BD Bioscience, New Jersey, USA), LIMP-2 (WB, 1:250; IF, 1:2000; kindly provided by J. Blanz), MAC-3 (1:400; 553322 BD Bioscience), Na_v1.6 (1:500; ASC-009 Alomone labs, Israel), NF155 (1:1000; kindly provided by P. Brophy), P0 (1:1000 [Archelos et al., 1993 (link)]); P2 (1:500; sc-49304 Santa Cruz), PLP (1:5000 [Jung et al., 1996 (link)]), PMP22 (1:1000; SAB4502217 Sigma), PMP70 (1:600; ab3421 Abcam), RAB7 (1:300; R4779 Sigma), TAG-1 (1:500; kindly provided by E. Peles), ßiv spectrin (1:400; kindly provided by M. Rasband), α-Tubulin (1:1000; T 5168 Sigma), III β-Tubulin (1:1000; Covance, New Jersey, USA).

Total liver protein lysates were obtained and separated in 10% sodium dodecyl sulfate-poly-acrylamide gel electrophoresis (SDS-PAGE). Gels were then blotted onto PVDF Amersham Hybond-P membranes (GE Health-care, Buckinghamshire, UK) and incubated with their corresponding antibodies (A2228, A9917, C0979 and S2147 from Sigma–Aldrich, St. Louis, Missouri, USA; Ab59546 from Abcam, Cambridge, UK; sc-2004, sc-2490 and sc-32886 from Santa Cruz Biotechnology, Dallas, Texas, USA). β-actin was used as loading control. Blots were developed by enhanced chemiluminescence using an Amersham ECL Plus Western Blotting Detection Reagent (GE Health-care, Buckinghamshire, UK) according to manufacturer’s instructions.

Proteins were isolated from microdissected formalin‐fixed, paraffin‐embedded (FFPE) cerebellum sections using the Qproteome FFPE Tissue kit (Qiagen, Valencia, CA) according to the manufacturers' instructions. Subsequent Western blotting of protein lysates and densitometric analysis were carried out as previously described.29 Primary antibodies used were anti‐β actin (ab8227, 1:5,000, Abcam), anti‐catalase (C0979, 1:2,000, Sigma‐Aldrich), anti‐human frataxin (ab110328, 1:2,000, Abcam), anti‐frataxin (ab113691, 1:2,000, Abcam), anti–glutathione peroxidase‐1 (ab22604, 1:1,000, Abcam), anti‐NeuN (ab177487, 1:5,000, Abcam), anti–nuclear factor (erythroid‐derived 2)‐like 2 (Nrf2; sc‐722, 1:3,000; Santa Cruz Biotechnology, Santa Cruz, CA), anti‐peroxisome proliferator‐activated receptor gamma coactivator 1‐alpha (PGC‐1alpha) (sc‐13067, 1:3,000, Santa Cruz Biotechnology), anti‐SOD1 (ab16831, 1:5,000, Abcam), and anti‐SOD2 (ab16956, 1:5,000, Abcam). (Of note, increases in total frataxin are likely to be underestimated when compared to human frataxin due to differences in antibody (ab113691, Abcam) species reactivity; the human protein reactivity of the antibody is approximately 50% of mouse protein reactivity at the same concentration. Commercial mouse‐specific frataxin antibodies are not available.)