Fig. 3: Nitration of aSyn does not impair the capacity of pS129 antibodies to detect pS129 fibrils.

a Preparation of aSyn fibrils carrying both phosphorylation and nitration modifications. aSyn fibrils were first site-specifically phosphorylated at S129 residue by being incubated with PLK3. Afterwards, TNM was added to induce the nitration of the phosphorylated aSyn fibrils. b The capacity of aSyn fibrils site-specifically phosphorylated at S129 and/or nitrated to be detected by the pS129 commercial antibodies (pSyn#64, MJF-R13, 81A, and EP1536Y) or the LASH-EGT homemade pS129 antibody was assessed by slot blot (c) and WB (d) analyses. a, b were created using BioRender.com. c, d Thirty-six nanograms of aSyn fibrils (PFFs) that were unmodified (WT), phosphorylated (P), nitrated (N), or nitrated/phosphorylated (N/P) were detected by slot blot (c) and WB (d) using pS129 antibodies (top panel) in combination with total aSyn antibodies (1–20 and 34–45, bottom panel). All blots were derived from the same experiment and were processed in parallel.