Figure 6.

UA induces K⁴⁸-linked ubiquitination (Ub) of Mcl-1. RASFs were pretreated with MG-132 (10 μM) for 4 h, followed by UA (10 μM) treatment for 24 h. The cells were lysed in denatured conditions (1% SDS and boiling) and then diluted before IP with anti-Mcl-1 antibody. Immunoblotting was performed with anti-Ub, K⁴⁸ Ub, K⁶³ Ub, or Mcl-1 antibodies. The experiment was performed twice.