Fig 1: Effect of OGD on calpain activation and on Rpn6, Rpn10 and Rpt3 total protein levels in cultured cerebrocortical neurons. Cultured cerebrocortical neurons were subjected to OGD for 1.5 h and were further incubated in culture conditioned medium for 4 h. When appropriate, cerebrocortical neurons were incubated with 100 µM APV or 50 µM MDL28170. The inhibitors were added 30 min prior and during OGD, and were also present during the post-incubation period. (A) Spectrin full-length (280 kDa) and SBDPs (150 and 145 kDa) were analyzed by western blot using an antibody recognizing both the N- and C-terminal regions of the protein. The total abundance of (B) Rpn10, (C) Rpt3 and (D) Rpn6 proteins levels were also evaluated by immunoblot. β-tubulin was used as loading control. Control (Sham without MDL or APV) protein levels of spectrin were set to 100%. The results are the average ± SEM of 3 independent experiments. Statistical analysis was performed by one-way ANOVA, followed by the Bonferroni´s multiple comparison test, comparing all the conditions with the respective control (ns, p > 0.5; ++++p˂ 0.0001; *p˂ 0.5; ***p˂ 0.001 ****p˂ 0.0001)