Fig. 6.

L-type calcium channel blocker reversed PrP (106–126)-mediated reduction in AMPK activity and increased autophagy flux in neuronal cells. a The primary neuronal cells were incubated with L651,582 for 1 h and then exposed to PrP (106–126) (100 μM) for 6 h. The treated cells were evaluated for p-AMPK by western blot analysis, indicate that L651,582 increased p-AMPK. b The bar graph represents the mean p-AMPK levels. c Immunocytochemistry for the p-AMPK was performed in SK-N-SH cells. d The bar graph represents the mean green fluorescence intensity. e Primary neuronal cells were incubated with isradipine and L651,582 for 1 h and then treated with PrP (106–126) (100 μM) for 6 h. The cells were evaluated for P62 and LC3B expression by western blot analysis, indicate that isradipine and L651,582 increased p62. f The bar graph represents the mean p62 and LC3B-II levels. g SK-N-SH cells were treated with GFP-LC3B virus for over 18 h and then exposed to PrP (106–126) for 6 h with isradipine or L651,582. Cell nuclei were stained with DAPI (blue) and evaluated by confocal microscopy. The cells were treated with negative and positive control reagent (CQ) at the same time. Western blot, immunocytochemistry, and GFP-LC3B puncta formation assay results represent at least three independent experiments. Data are expressed as the mean ± SEM. * p < 0.05, ** p < 0.01; significant differences between each treatment group (Welch’s T-test). PrP, Prion peptide (106–126); isra, isradipine; adj.volume, adjustment of volume (band volume minus background volume)