Supplementary Figure 4: Immunocytochemical analysis of AZ proteins in Munc13-1 or RIM1 knockdown synapses.
From: Synaptic weight set by Munc13-1 supramolecular assemblies
a Representative immunofluorescence images of major AZ proteins, including Munc13-1, RIM1, bassoon, piccolo, and ERC1b/2, in control, Munc13-1 shRNA lentivirus-infected neurons and RIM1 shRNA lentivirus-infected neurons. Scale bar, 5 µm. Images are representative of 4 experiments. b A summary graph showing the average immunofluorescence intensity of AZ proteins at synapses (n = 60 synapses each from 4 culture preparations). Lentiviral expression of Munc13-1 shRNA reduces the immunofluorescence intensity of Munc13-1 to 11% of control. Lentiviral expression of RIM1 shRNA reduces the immunofluorescence intensity of RIM1 and of Munc13-1 to 18% and 48% of control, respectively. Data shown are mean ± SEM. Two-tailed Wilcoxon rank sum test was used for statistical analysis. Munc13-1: P = 1.7×10−17(W = 3423) for control v.s. shMunc13-1, P = 0.000013 (W = 2632) for control v.s. shRIM1; RIM1: P = 0.14 (W = 2083.5) for control v.s. shMunc13-1, P = 1.1×10−17 (W = 3431) for control v.s. shRIM1; Bassoon: P = 0.15 (W = 1525) for control v.s. shMunc13-1, P = 0.23 (W = 1570) for control v.s. shRIM1; Piccolo: P = 0.59 (W = 1904) for control v.s. shMunc13-1, P = 0.026 (W = 2226) for control v.s. shRIM1; Erc1b/2: P = 0.59 (W = 1698) for control v.s. shMunc13-1, P = 0.16 (W = 1530) for control v.s. shRIM1. **P < 0.001; †0.01< P < 0.05; n.s., not significant, P > 0.05.
