Fig. 3
From: Two di-leucine motifs regulate trafficking and function of mouse ASIC2a
AAD and DAA mutants exhibit increased dendritic trafficking. Organotypic ASIC2−/− hippocampal slices were transfected with HA-tagged ASIC2a WT, AAD, or DAA mutant together with a membrane-targeted Lck-GFP. Localization of ASIC2a was detected using an anti-HA antibody. a Top: Representative images showing the overall distribution of ASIC2a in hippocampal pyrmaidal neurons. Bottom: Quantification of ASIC2a dendrite:cell body ratio. Two lines illustrate the position used for quantification: at mid-apical dendrite and cell body. The ratio is calculated as: (dendritic ASIC2a/cell body ASIC2a)/(dendritic GFP/cell body GFP). The cell body level calibrates for changes in expression while the ratio of GFP calibrates for changes in volume. p values were from one way ANOVA. b. Top: High magnification images showing ASIC2a localization in segments of mid-apical dendrites. Bottom: Quantification of ASIC2a spine:shaft ratio. To calibrate for changes in expression and volume, for a given spine, the ratio is calculated as: (spine head ASIC2a/shaft ASIC2a)/(spine head GFP/shaft GFP). p values were from ANOVA followed by Turkey’s HSD test. N on the bars represent total number of neurons (a) or spines (b) quantified