Formation of detergent/salt-insoluble ubiquitinated-protein aggregates following OGD/reperfusion, and the inhibition of 1400W.
A. Protein smears ranging from 37 to 250 kDa were considered to be of ubiquitinated proteins. Increased levels of detergent/salt-insoluble ubiquitinated-protein aggregates were observed in pellet fractions under OGD/reperfusion. This change was more pronounced in OGD 8 h/reperfusion 24 h group. 1400W inhibited the formation of ubiquitinated-protein aggregates in the insoluble pellet fraction, with the maximum effect seen at the concentration of 50 μM. B. The ubiquitinated-protein aggregates were quantified using average intensity of protein smear, using β-actin as an internal control. Data from three independent experiments were presented as mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001 by one-way ANOVA (analysis of variance) with Tukey’s post-hoc test.