Figure 5.
Immunoblot analysis for protein-bound nitrotyrosine in familial and sporadic ALS patients. Soluble fractions (A) or neurofilament-enriched fractions (B) from sporadic ALS patients (lanes 1–3), control individuals (lanes 4–6), and familial ALS patients with SOD1 mutation A4V (lanes 7 and 8) were immunoblotted with nitrotyrosine antibody 1A6 (Top). (Middle) Parallel immunoblots with the same antibody but in the presence of 10 mM 3-nitro-l-tyrosine. (Bottom in A) The same whole spinal cord extracts stained with Coomassie blue. (Bottom in B) The blot of the neurofilament fractions in B Top was stripped and reprobed with an antibody to NF-L. Lanes 9, an in vitro nitrated neurofilament sample. The A4V samples in lanes 7 and 8 were from 63- and 42-year-old patients 6 or 17 hr post mortem. The SALS samples in lanes 1, 2, and 3 are from 81-, 62-, and 61-year-old patients 7, 14, and 13 hr post mortem.