Research Paper Volume 8, Issue 7 pp 1470—1484

Figure 3. NPY receptor antagonists inhibit the stimulatory effect of caloric restriction on autophagy in rat cortical neurons. Primary rat cortical neurons were incubated with NPY Y₁ receptor antagonist BIBP3226 (Y₁ant, 1 μM), NPY Y₂ receptor antagonist BIIE0246 (Y₂ant, 1 μM) or NPY Y₅ receptor antagonist L152,800 (Y₅ant, 1 μM), 30 min before caloric restriction medium (CR) for 6 h. Whole cell extracts were assayed for LC3B-II (A), SQSTM1 (B) and β-tubulin (loading control) immunoreactivity by Western blotting analysis, as described in Materials and Methods. Representative Western blots for each protein are presented above each respective graph. The results represent the mean ± SEM of, at least, five independents experiments, and are expressed as percentage of control. *p<0.05 and ***p<0.001, significantly different compared to control; ^#p<0.05, ^##p<0.01 and ^###p<0.001, significantly different from caloric restriction, as determined by ANOVA, followed by Bonferroni’s post test.