Fig 2. TH ubiquitination.

PC12 cells were incubated for 30 min in the absence (basal) or presence of lactacystin (Lacta) (1 μmol/L). Ub-TH was immunoprecipitated with an ubiquitinated protein enrichment kit (IP: anti-Ub). Proteins were resolved in a 4–12% SDS-PAGE and Ub-TH protein level assessed by Western-blot as described under Methods. The same membrane was stripped and reprobed with an antibody against ubiquitin (Ub) to further corroborate that Ub-proteins were isolated with the kit. Three separate experiments were performed. A representative Western blot immunoblotted with antibodies directed against TH or Ub is shown.