Fig. 5: BAG5 knockdown partially rescues Mcl-1 degradation.
From: Bcl-2-associated athanogene 5 (BAG5) regulates Parkin-dependent mitophagy and cell death
a Western blot illustrating the changes in endogenous Mcl-1, BAG5, and exogenous myc-Parkin elicited by siRNA-mediated BAG5 knockdown (siBAG5) vs. Control (siNTC) with or without CCCP treatment in SH-SY5Y cells. b Quantification of Mcl-1 presence in the western blot. Mcl-1 intensity normalized to Actin and expressed as a ratio of the intensity found in the siBAG5 lane vs. siNTC lane. Data obtained from three independent experiments, normalized to siNTC condition and compared using an unpaired t test (*p < 0.05). Columns represent mean ± SEM. c SH-SY5Y cells overexpressing GFP-BAG5 or GFP were transfected with HA-ubiquitin plus siRNA targeting Parkin (siParkin) or control siRNA and then treated with 50 μm CCCP for 18 h. Ubiquitinated proteins were immunoprecipitated with anti-HA antibodies and analyzed by western blot. *Indicates previously probed band from panel below. d Quantification of the level of immunoprecipitated HA-ubiquitinated Mcl-1 (HA-Ub Mcl-1) relative to Mcl-1 protein levels in the inputs. Data obtained from three independent experiments and statistical analysis was done using one-way ANOVA followed by Tukey post hoc testing. Columns represent mean ± SEM.
