A longer time spent in the correct quadrant is indicative of a learning effect wherein the mice remember the previous location of the platform and spend an increased period of time looking for it, whilst a shorter time in this quadrant indicates a memory deficit. displaying immunostaining in the neuropil and neurons respectively. (H, I) Confocal images with the 9E4 antibody in non-tg showing no specific labeling and immunostaining in the neuropil and neurons in -syn tg. (J, K) FL -syn immunoreactivity in non-tg and -syn tg, respectively. (L, M) CC -syn immunoreactivity in non-tg and -syn tg, respectively). N?=? 3 per group, 6 months of age. Scale bar (B-M) ?=? 30M.(TIF) pone.0019338.s001.tif (1.0M) GUID:?BEC2A24D-3BCD-435E-B959-CCE84F4BFC00 Figure S2: Effects of passive immunization with a C-terminal -syn antibody on tau. Levels of total and PHF-tau were examined by immunohistochemistry and immunoblot analysis to asses the effects of passive immunization with 9E4. (A, B) Representative brightfield images of total tau in the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively. (C, D) Representative brightfield images of total tau in the frontal cortex of IgG1- and 9E4-treated -syn tg mice, respectively. (E) Quantitative analysis of total tau levels in the frontal cortex of IgG1 and 9E4-treated non-tg and -syn tg mice. (F, G) Representative brightfield images of PHF-tau in the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively. (H, I) Representative brightfield images of PHF-tau in the frontal cortex of IgG1- and 9E4-treated -syn tg mice, respectively. (J) Quantitative analysis of PHF-tau levels in the frontal cortex of IgG1 and 9E4-treated non-tg and -syn tg mice. (K) Immunoblot of levels of total and PHF-tau in the frontal cortex of IgG1 and 9E4 treated non-tg and -syn tg mice. (L) Analysis of total tau levels in the frontal cortex of IgG1 and 9E4-treated non-tg and -syn tg mice as determined by immunoblot. (M) Analysis of PHF-tau levels in the frontal cortex of IgG1 and 9E4-treated non-tg and -syn tg mice as Almitrine mesylate determined by immunoblot. Scale bar ?=? 40uM. Error bars represent mean SEM. (*) Indicates p<0.05, when comparing -syn tg immunized with IgG1 or 9E4 to IgG1-treated non-tg mice by one-way ANOVA with post hoc Dunnett's.(TIF) pone.0019338.s002.tif (2.5M) GUID:?B282893D-E3B2-4E64-A2F6-C0127BEEA8E2 Figure S3: Effects of passive immunization with a C-terminal -syn antibody on vasculature or markers of glial cell reactivity. (A, B) Representative brightfield images of the endothelial cells marker Zo-1 immunoreactivity in Almitrine mesylate the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively (arrows indicate location of blood vessels). (C, D) Representative brightfield images of Zo-1 immunoreactivity in the frontal cortex of IgG1- and 9E4-treated -syn tg mice, respectively Almitrine mesylate (arrows indicate location of blood vessels). (E) Analysis of % of Zo-1 immunoreactive neuropil in the frontal cortex of IgG1- and 9E4-treated non-tg and -syn tg mice. (F, G) Representative brightfield images of the microglial marker Iba-1 immunoreactivity in the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively. (G, H) Representative brightfield Almitrine mesylate images of Iba-1 immunoreactivity in the frontal cortex of IgG1- and 9E4-treated -syn tg mice, respectively. (E) Analysis of Iba-1 immunoreactivity in the frontal cortex of IgG1- and 9E4-treated non-tg and -syn tg mice. (F, G) Representative brightfield images of the microglial marker Iba-1 immunoreactivity in the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively. (K, L) Representative brightfield images of the astroglial cell marker GFAP immunoreactivity in the frontal cortex of IgG1- and 9E4-treated non-tg mice, respectively (arrows indicate location of blood vessels). (M, N) Representative brightfield images of GFAP immunoreactivity in the frontal cortex of IgG1- and 9E4-treated -syn tg mice, respectively (arrows indicate location of blood vessels). (E) Analysis of GFAP immunoreactivity in the frontal cortex of IgG1- and 9E4-treated non-tg and -syn tg mice. Scale bar (A-D) ?=? 80uM, (F-N) ?=? 50M. Error bars represent mean SEM. (*) indicates p<0.05, Rabbit polyclonal to IWS1 when comparing IgG1-immunized -syn tg mice to IgG1-imunized non-tg mice by one-way ANOVA with post hoc Dunnett’s..