Fig. 6: CK2, p-CaM, and SK3 protein expression are increased in 3xTg VTA DA neurons.

a Workflow for cell segmentation analysis following immunohistochemistry. Confocal z-stack images were acquired, then the brightest slice of the channel of interest was chosen for analysis. DA neurons were segmented on TH staining from the same plane using the Cellpose ‘cyto v3’ model following upscaling (see Methods). Intensities within each segmented cell were calculated, quantified as average intensity across single cells, and plotted as cumulative probabilities. b Staining for CK2 indicated a slight rightward shift in the cumulative probability curve (c) of casein kinase 2 (CK2) intensity in tyrosine hydroxylase (TH) positive cells from 3xTg mice (Kolmogorov–Smirnov test, D = 0.1183, p < 0.0001, n = 2353 WT and 2187 3xTg cells, 6 mice). d Staining for phosphorylated calmodulin (p-CaM) indicated a moderate rightward shift in the cumulative probability curve (e Kolmogorov–Smirnov test, D = 0.1202, p < 0.0001, n = 2612 WT and 2193 3xTg neurons, 6 mice). f SK channel upregulation in 3xTg mice was indicated by a strong rightward shift in the cumulative probability of small conductance calcium-activated potassium channel 3 (SK3) intensity in single cells (g Kolmogorov–Smirnov test, D = 0.2284, p < 0.0001, n = 2914 WT and 3165 3xTg neurons, 6 mice). Red scale bars = 200 µm. Source data are provided as a Source Data file.