Subcellular localization of native rEag1 and rEag2 channels in young and mature hippocampal neurons in culture. Dissociated hippocampal neurons at DIV3, DIV7, and DIV12 were immunostained with the anti-rEag1 (A, B) or the anti-rEag2 (C, D) antibodies (shown in green; left panels), followed by counterstaining with the antibody for the dendritic marker MAP2 or the axonal marker tau (shown in red; middle panels). Merged images are shown in the right panels. (A) rEag1 immunoreactivities were localized in cell bodies, as well as in MAP2-positive and MAP2-negative (arrows) processes. (B) rEag1 immunoreactivities were present in the axonal compartment that was clearly defined by the immunofluorescence signal of tau (arrows). For DIV12 neurons in both (A) and (B), note the presence of punctate rEag1 staining patterns throughout proximal and distal neurites. (C, D) rEag2 immunoreactivities were present in both MAP2-positive and tau-positive processes. No significant rEag2 puncta were observed. Scale bar, 25 μm.