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. 2018 Apr 30;11:142. doi: 10.3389/fnmol.2018.00142

Figure 2.

Figure 2

Cdh2 on 293TA cells provides favorable surface for neurite branching of cocultured cortical neurons. (A–D,A′–D′) Mouse cortical neurons were cultured with 293T derivatives and stained with anti-synapsin I (red) and phalloidin (green). (A) Cortical neurites frequently branch on 293TA cells as shown in the high-power picture (bottom; region indicated by blue arrow in top panel). (B) Neurites generally grew unbranched near the edge of 293NC cells (C) Neurites branched on Cdh2-overexpressing 293NC (D) Neurites failed to branch on neural cell adhesion molecule (NCAM)-overexpressing 293NC cells. Bar, 25 μm for (A–D), and 5 μm for high-power images (bottom). (E) Percent of neurons exhibiting branching at sites of apposition with 293 cells (n = 20 neurons per condition). Co-expression of NLGN1 had no effect on branching in the presence or absence of Cdh2. (F,G) Quantification of calcium-dependent (F) and independent (G) cell aggregation assays (Mean ± SEM, n = 10). Significance of differences at 60 min and 120 min, p < 0.005 by Student’s t-test between 293NC and 293NC+NCAM in (F,G); p < 0.05 between 293NC and 293NC+Cdh2EC in (F); p < 0.05 between 293NC+Cdh2 and 293 + Cdh2EC in (F,G); p > 0.1 between 293NC and 293NC+Cdh2EC in (G). Data on 293NC and 293NC+Cdh2 cells replotted from Figure 1. 293NC cells transfected with mouse NCAM showed calcium-independent cell aggregation. 293NC cells transfected with chicken Cdh2EC showed calcium-dependent cell aggregation.