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. 2016 Mar 24;7:11080. doi: 10.1038/ncomms11080

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(a) Representative confocal images from organotypic hippocampal slices transfected with pSuper plasmid containing BRAG1-siRNA and expressing GFP (to visualize the transfected cells) and immunostained 2 days later for BRAG1 (red signal) under permeabilized conditions. Note, the cell transfected with the BRAG1-siRNA (green cell) has low levels of endogenous BRAG1 (red signal). Scale bars, 10 μm. (b) Representative line plot analysis of BRAG1 (red signal) and GFP (green signal). CB1 is a cell body of non-transfected neuron; CB2 is that of a transfected neuron. (c) Normalized intensity from the line plots for endogenous BRAG1 for the three different RNAi used; RNAi significantly (P≤0.05) decreased BRAG1 levels. RNAi #1: n=14, P=0.0112; RNAi #2: n=7, P=0.0132; RNAi #3: n=10, P=0.0417. (d,e) Simultaneous whole-cell double recordings from nearby pairs of untransfected (control) neurons and those transfected with either BRAG1-siRNA#2 (d) or BRAG1-siRNA#3 (e). Left graph: comparisons of evoked AMPAR-mediated responses. Right graph: simultaneous recordings of evoked NMDAR-mediated responses (P=0.21). (f) RNAi #3 was co-expressed with RNAi-resistant WT BRAG1 to restore BRAG1 levels. Data represent average AMPAR EPSCs (left; n=10, P=0.15), NMDAR EPSCs (middle; n=8, P=0.51) and AMPA/NMDA ratios (right; n=8, P=0.60). * Indicates significance (P≤0.05).