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. 2022 Apr 8;11:e75707. doi: 10.7554/eLife.75707

Figure 2. Design and characterization of Syngap1 IRES-TdTomato knock-in mice.

(A) IRES-TdTomato insertion site in relation to α1 and α2 open-reading frames. (B) Genotype ratios and survival probability following heterozygous breeding. (C) Representative western blots showing expression levels of total SynGAP and individual isoforms. Quantification of forebrain expression levels measured by western blot analysis. Relative intensity of bands normalized to total protein signal. Only α1 signal is significantly changed. ANOVA with Tukey’s multiple comparisons test, F(2, 14) = 24.86, n = 5, p < 0.0001 (D) Quantification of total distance traveled in open field test in adult WT or Td/Td mice. Unpaired t-test t(4)=10.42. Note that very few homozygous Td/Td mouse survived through adulthood. (E) Quantification of total distance traveled in open field test in adult WT or +/Td mice. Unpaired t-test t(18)=9.007 (F) Latency of event onset was measured as the time taken to 1st clonus (seizure onset). Unpaired t-test: t(18)=2.588. (G) Percent freezing in remote contextual fear memory paradigm. Unpaired t-test: t(41)=2.49 (H) Plots demonstrating latency to find platform across days in Morris Water Maze training. Linear mixed model for repeated measures. n = 9–12, +/+ vs + /Td, p = 0.3.

Figure 2—source data 1. Representative blots and total protein profiles.

Figure 2.

Figure 2—figure supplement 1. mRNA and protein isoform expression in Syngap1 mouse models.

Figure 2—figure supplement 1.

(A) Representative western blots demonstrating total SynGAP and isoform expression level in forebrain lysates from Syngap1+/+ and Syngap1+/- +/-. Relative intensity of bands normalized to total protein signal. Statistical significance is determined by unpaired t-test. Total: t(4)=5.403, α1: t(4)=9.044, α2: t(4)=4.473, β: t(4)=3.931 (B) Syngap1 exon usage in +/+, +/Td, and Td/Td mice. (C) Representative western blots showing expression levels of total SynGAP and individual isoforms at PND110 from in +/+ and +/Td mice. Unpaired t-test. Total: t(6)=1.784, α1: t(6)=3.351, α2: t(6)=5.678, β: t(6)=4.425.
Figure 2—figure supplement 1—source data 1. Representative blots and total protein profiles.