Figure 3.
Evaluating the variability of presynaptic induction using software-based analysis of synaptic vesicle (SV) clusters induced by LRRTM2. (A) Raw confocal images (20×) of GFP-expressing neurons cultured with HEK293T cells transfected with mCherry alone (top row) or mCherry and LRRTM2 (bottom row) and immunostained against synaptobrevin. The overlap region map and corresponding SV clusters (black) is displayed on the right. Scale bar, 20 μm. (B) Mean percentage of axon-HEK293T overlap regions containing SV clusters (SV cluster frequency) from six different fields, across three independent experiments. HEK293T cells express mCherry (control) or mCherry together with LRRTM2. p < 0.001, ANOVA. No significant difference (p = 0.78) between the three independent neuronal preparations. (C) Fraction of area of overlap region occupied by SV clusters (SV cluster density). p < 0.05, ANOVA. No significant difference (p = 0.44) between the three independent neuronal preparations.