Fig. 4. Localization of Necl-1 in cocultured neurons and glia cells. Immunofluorescence images of mouse hippocampal cocultured neurons and glia cells at 24 days in vitro. (A) Contact sites of neurons. Cells were double stained with anti-Necl-1 pAb and anti-synaptophysin mAb. Aa, Necl-1 (red); Ab, synaptophysin (green); Ac, merge. (B) Contact sites of glia cells. Cells were double stained with anti-Necl-1 pAb and anti-GFAP mAb, a marker for glia cells. Ba, Necl-1 (red); Bb, GFAP (green); Bc, merge. Arrows indicate contact sites of glia cells. (C,D) Contact sites between a neuron and a glia cell. Cells were double-stained with anti-Necl-1 pAb and anti-GFAP mAb, a marker for glia cells. Ca and Da, Necl-1 (red); Cb and Db, GFAP (green); Cc and Dc, merge. Brackets indicate contact sites between a neuron and a glia cell. Inset in C, images at a higher magnification. Bars, 10 µm. The results shown are representative of three independent experiments.