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. 2014 Jul 9;9(7):e101770. doi: 10.1371/journal.pone.0101770

Figure 6. Loss of α-actinin-2 prevents synapse formation.

Figure 6

A) Actively firing pre-synaptic boutons do not synapse with spines on neurons lacking α-actinin-2. Hippocampal neurons were co-transfected at DIV 6 with GFP and either pSUPER or pSUPER-α-actinin-2, treated with FM4-64 for 5 min on DIV 19 and observed live. Arrows mark FM4-64 juxtaposition to spines or the dendrite. The fraction of spines juxtaposed to FM4-64 is reduced in neurons lacking α-actinin-2 (see results). For each condition, 14 neurons from 2 separate cultures were analyzed. B) Loss of α-actinin-2 prevents synapse formation with excitatory axon boutons. Hippocampal neurons were co-transfected at DIV 6 with GFP and either pSUPER or pSUPER-α-actinin-2, fixed on DIV 21, and immunostained for VGLUT1. Arrows mark VGLUT1 juxtaposition to spines or the dendrite. The fraction of spines juxtaposed to VGLUT1 is reduced in neurons lacking α-actinin-2 (see results). For each condition 23–26 neurons from 2 separate cultures were analyzed.