Figure 2

Expression of neurocan and NCAM in postnatal and adult brain. (A) Immunoblot of total brain extracts for neurocan, NCAM, PSA-NCAM, and GAPDH (loading control). (B) Co-localization of tdTomato puncta (red) with the presynaptic marker GAD65 (green) around a neuronal soma (blue). Scale bar = 5 μm. (C) Immunofluorescent staining of neurocan (green) around PV⁺ tdTomato-labeled interneurons (red) in mouse frontal cortex at P28 and P60. Scale bar = 50 μm. (D) Confocal images of neurocan (green) around perisomatic synaptic puncta (red) onto MATH-2 expressing pyramidal neurons (blue nucleus, outlined with white border). A magnified inset is indicated with a white box. Scale bars = 2.5 μm. (E) Electron micrograph of immunogold labeling for neurocan along the membrane of a neuronal soma (black arrows point to examples). Scale bar = 1 μm. (F) Electron micrograph of an inhibitory axon terminal onto a neuronal soma. Neurocan immunogold labeling is indicated with black arrows, and the axon terminal (AT) is labeled with a white arrowhead. Cytoplasm and nucleus of the soma are identified with text. Scale bar = 1 μm. (G) Verification of neurocan antibody specificity by immunostaining of COS7 cells transfected with Neurocan-AP (+control) or AP (−control), and a no primary antibody control. Scale bar = 50 μm.(H) Immunofluorescent localization of NCAM and neurocan in GABA+ and GABA− cortical neuron cultures. Confocal images of GABA (green), NCAM (blue), and neurocan (red) in cortical neuron cultures. White arrow indicates a GABA-negative neuron positive for NCAM and neurocan, and the green neuron represents a GABAergic interneuron. Scale bar = 10 μm.