Figure S1. Evaluation of Munc18-1A Protein Expression in the Three Transgenic Lines Of

Figure S1. Evaluation of munc18-1a protein expression in the three transgenic lines of mice (a) Increased immunodensity of munc18-1a protein in brain cortex of transgenic mice from line 3 (L3) overexpressing munc18-1a (unpaired t test **P<0.005 versus wild type (WT) controls). (b) Increased immunodensity of munc18-1a protein in brain cortex of transgenic mice from line 7 (L7) overexpressing munc18-1a (unpaired t test *P<0.05 versus WT controls). (c) No significant changes were found in munc18-1a expression in brain cortex of transgenic mice from line 8 (L8) when comparing to their WT controls. Data bars are mean ± s.e.m. values; the number of animals is indicated inside the plot bars. After initial characterization, the line 7 (termed Munc18-OE) was selected for further studies of munc18-1a overexpressing mouse phenotype.

Figure S2. Presynaptic proteins (a) No significant differences were found in SNAP25, synaptotagmin and synaptophysin between Munc18-OE mice and their wild-type controls. Representative immunoblots of SNAP25, synaptotagmin, synaptophysin and beta-actin proteins in Munc18-OE and WT control mice. (b) No significant differences were found in syntaxin-1A and SNARE complex between Munc18-OE mice and WT controls. Representative immunoblots of syntaxin-1A, SNARE complex (antibody for syntaxin 1-A) and beta actin proteins. (c) No significant changes were observed in synaptobrevin protein in Munc18-OE mice when compared to WT controls. Representative immunoblots of synaptobrevin, SNARE complex (antibody for synaptobrevin) and beta actin proteins. Data bars of inmunoblots are mean (percentage over control) ± s.e.m.; the number of animals is indicated in parentheses.

Figure S3. Complementary behavior (a) No significant differences were found in holeboard test, as shows the number of nosepokes, between Munc18-OE and their wild-type (WT) controls. (b) No significant differences were found between Munc18-OE and WT control mice in the rotarod test for the evaluation of motor coordination. Bars represent time in seconds in the rotarod apparatus. (c) No changes were observed in the forced swim test in Munc18-OE mice when compared to their WT controls. Bars represent time of immobility in seconds. (d) Schematic representation of water maze test for evaluation of spatial memory (up). No differences were found in time spent in each quadrant in probe or reversal test (down). Bars represent percentage of time in each quadrant of the maze.

Data bars are mean ± s.e.m. values; the number of animals is indicated in parentheses or inside the plot bars.

FigureS4. Glutamate receptors (a) No significant changes were found in mGlu2 and mGlu3 metabotropic glutamate receptors between Munc18-OE and their wild-type controls. Representative immunoblots in WT controls and Munc18-OE mice. (b) No significant changes were shown in AMPAR1 and AMPAR2/3/4 receptor subunit densities of glutamate AMPA receptors between Munc18-OE and WT controls. Representative immunoblots in WT controls and Munc18-OE mice. (c) No changes were observed in NR1 and NR2A subunit densities of NMDA receptors in transgenic Munc 18-OE mice when compared to WTcontrols (d) Protein expression of PSD95 and Src did not differ between Munc18-OE and WT control mice. Data bars of immunoblots are mean (percentage over controls) ± s.e.m.; the number of animals is indicated in parentheses.