Tissue sections were cut from blocks of formalin-fixed paraffin embedded ALS tissue, obtained from Dr. John Ravits’ bank collection (control patient #83 and #65). The spinal cord sections of patients harboring TDP-43 mutations (Pt #284 and #203) as well as control #150 were obtained from the MRC London Neurodegenerative Diseases Brain Bank (King’s College London) (see table below). Seven µm-thick tissue sections were stained with an antibody against RanGAP1 (ab92360) from Abcam (Cambridge, MA, USA). Slides were stained on a Ventana Discovery Ultra (Ventana Medical Systems, Tucson, AZ, USA). Antigen retrieval was performed using Tris–EDTA based cell conditioning solution (CC1) for 40 min at 95 °C. The primary antibody was incubated on the sections at 1:300 dilution for 32 min at 37 °C followed by UltraMap (Ventana) and DAB detection. Slides were rinsed, dehydrated through alcohol and xylene, and coverslipped. Imaging was performed on Nanozoomer slide scanner (Hamamatsu) at the UCSD microscopy core.
| Patient # | Age at death (years) | Sex | Diagnosis | Genotype | Post-mortem delay + interval (h) | Reference |
|---|---|---|---|---|---|---|
| 65 | 82 | M | NORMAL | N/A | 4* | |
| 83 | 63 | F | NORMAL | N/A | 4* | |
| A150/01 | 40 | M | NORMAL | N/A | 40 | |
| A284/13 | 57 | M | ALS | TARDBP M337V | 74 | [60 (link)] |
| A203/12 | 23 | F | ALS | TARDBP Y374X + FUS P525L | 125 | [32 (link)] |
* Post-mortem interval only (post-mortem delay not available)
Free full text: Click here
