Double immunofluorescence of PMCA2 or PMCA3, and synaptophysin in different brain regions at P30 stage. Control slides without the primary antibody are shown to verify specificity of the fluorescence signals. PMCA2 and PMCA3 (red) colocalized with synaptophysin (green) in the neuropil (asterisks) and in cerebellar glomeruli (short arrow), confirming the presence of these pumps in synaptic areas (merge). DAPI staining (blue) was used to visualize nuclei. so, stratum oriens; pcl, pyramidal cell layer; sr, stratum radiatus; ml; molecular layer, Pcl; Purkinje cell layer, gl; granular layer. Scale bar: 25 μm.