Licor odyssey infrared imaging system

Imaging of brain sections and CHO cells was performed with a Leica-SP8 confocal microscope. Western blot data acquisition was conducted with LiCOR Odyssey Infrared imaging system (Model No. 9120) and Universal hood II (Bio-Rad, California, USA). Mean fluorescence values, cerebellum diameters, areas and cell quantifications were calculated with Fiji Version 1.51g software (Image J) (http://fiji.sc/Fiji) (W. Rasband, NIH, USA). See Supplementary experimental procedures for analysis of cerebellar sections.

HEK293 supernatant or purified CaMKII protein concentration was determined by the BCA method. Protein samples were mixed with 4× NativePAGE Sample Buffer (Thermo Fisher Scientific), NativePAGE 5% G-250 Sample (Thermo Fisher Scientific), and water for a final volume of 10 μl. The same amount of protein (20 μg for supernatant and 1 μg for purified protein) was loaded onto each lane, and Native-PAGE was performed on NativePAGE Novex 3 to 12% Bis-Tris gel. The gel was run with cathode buffer (1× NativePAGE Running Buffer, 1× NativePAGE Cathode Additive) and anode buffer (1× NativePAGE Running Buffer) at maximal 150 V for 2 h. The gel was stained with Coomassie Brilliant Blue R-250 (Bio-Rad) and scanned on a Li-Cor Odyssey infrared imaging system according to the manufacturer’s instructions.