Figure 6.

Validation of novel DUB targets of ataxin-3. (A) Immunoblot using antibody against HIF1a in whole cell lysate (input) and TUBEs1 enriched fractions from control and ataxin-3 shRNA THP-1 cells left unstimulated or stimulated for 60 min with MDP + PAM₃CSK₄. Immunoblot using antibodies against (B) PLD3 and (C) LAMTOR1 in whole cell lysates (input) and TUBEs1 enriched fractions from control shRNA and ataxin-3 shRNA THP-1 cells left unstimulated or stimulated for 60 min with MDP + PAM₃CSK₄. (D) Immunoblot using antibodies against GFP, HA, and β-actin in input and HA-immunoprecipitated lysates, in HEK293 cells where GFP and/or GFP-LAMTOR1 and/or HA-ataxin-3 were overexpressed. (E) Immunoblot using antibodies against ubiquitin, GFP, ataxin-3, and β-actin in input and isotype control (iso) or GFP immunoprecipitated lysates where ubiquitin and GFP-LAMTOR1 were overexpressed with or without HA-ataxin-3. All immunoblots are representative of at least two independent experiments.