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. 2020 Feb 28;11:136. doi: 10.3389/fphar.2020.00136

Figure 1.

Figure 1

Phenotype verification of Scn1a KO mice and the increase of SCN1A expression after liraglutide treatment in the brain of Scn1a KO mice. (A) PCR was used to identify wild-type and Scn1a KO heterozygous (F1) genotypes. (M: DNA marker). (B) RT-qPCR analysis of the Scn1a mRNA expression from brains of wild-type (WT) and Scn1a KO heterozygote (KO) mice (N = 24 mice per group; Student’s t-test, P < 0.001). (C) Representative western blotting showing SCN1A protein expression in brains of WT and Scn1a KO mice. (D) Summary graph of western blotting analysis demonstrating a significant reduction in the SCN1A protein in brains of Scn1a KO as compared to WT mice (N = 24 mice per group; Student’s t-test, P < 0.001). (E) Representative images of immunofluorescent staining with SCN1A (green), and DAPI (blue) in the ipsilateral side of the cortex of the WT and Scn1a KO mice. Scale bar: 75 μm for the full-scale images and 25 μm for the magnified images. (F) Summary graph of the average optical density of SCN1A fluorescent staining measured in images (E) demonstrating a profound increase of SCN1A in the cortex of Scn1a KO mice after liraglutide administration (N = 24 mice per group; Student’s t-test, P < 0.001). (G) RT-qPCR analysis of the SCN1A mRNA expression after liraglutide administration in brains of WT and Scn1a KO mice (N = 24 mice per group; One-way ANOVA, Student’s t-test). (H) Representative western blotting showing SCN1A protein expression in brains of the WT and Scn1a KO mice. (I) Summary graph of western blotting analysis demonstrating a dramatic increase in the SCN1A protein from brains of Scn1a KO after liraglutide administration (N = 24 mice per group; One-way ANOVA, Student’s t-test). The relative expression of SCN1A was normalized to reference controls Gapdh and β-actin in RT-qPCR and Western blotting, respectively. Data were presented as mean ± SD; ***, P < 0.001; ns, not significant. All experiments were performed in triplicate. WT, Wild-type; KO, Scn1a Knockout heterozygotes (F1); NS, Normal saline; Lira, Liraglutide.