Fig. 5

NeuN⁺ clusters have partial colocalization of MAP2 and synaptophysin, strong colocalization with Chromogranin A, presynaptic markers in their periphery, and nearby glia. A In the CA1 region of the hippocampus of 12-month old DPP6-KO mice, immunofluorescence shows MAP2 (red, 1:500, Millipore) and synaptophysin (green, 1:500, Sigma) partly colocalized in clusters of puncta; on the right is the high magnification image. Scale bar = 50 μm. B, c Immunofluorescence staining shows the novel puncta labeled with NeuN and the presynaptic markers vGluT1 (B; red, 1:500, Millipore) and synapsin1 (C red, 1:200, Novus), D The NeuN⁺ novel puncta colocalize very well with Chromogranin A (red, 1:500, Novus; Scale bar = 5 μm). E, F The novel puncta are surrounded by astrocytes labeled with marker GFAP (E red, 1:2000, DAKO) and (F) microglia labeled with marker Iba1 (green, 1:800, Wako). Scale bar = 50 μm. sp, stratum pyramidale; sr, stratum radiatum. Nuclei were counterstained with DAPI in blue