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. 2018 Jun 4;7:e36768. doi: 10.7554/eLife.36768

Figure 2. Trim28 adult knockout mice are viable and demonstrate reduced α-Syn and tau levels.

(A) Experimental approach to delete Trim28 from the adult body. (B) Kaplan-Meier survival curve of Adult knockout mice (UBC-CREERT2; Trim28flox/flox + TAM vs littermate controls). No significant differences in survival are observed. (C) qPCR analysis for Trim28 expression in midbrain (MB), cortex (CTX) and hippocampus (HIP) of Trim28 adult knockout mice and control littermates. (D) Western blot analysis of α-Syn, tau and Trim28 levels in hippocampi from Trim28 adult knockout mice and control littermates. In (B), n = 14–33 per group. In (C and D), n = 12–13 per group.

Figure 2.

Figure 2—figure supplement 1. Perinatal suppression of Trim28 in the brain is safe and decreases α-Syn and tau levels.

Figure 2—figure supplement 1.

(A) Approach to deplete Trim28 in the postnatal brain via RNAi. (B) Left panel demonstrates widespread expression of AAV across the brain. Epifluorescence of YFP expression in a representative 10-week-old mouse demonstrating widespread expression of the AAV throughout various brain regions. Brain regions depicted: (i) whole forebrain; (ii) caudal cortex; (iii) caudal cortex (zoom); (iv) hippocampus; (v) hippocampal CA1 region (zoom); (vi) Purkinje cells; and (vii) brainstem). Right panel denotes evidence of Trim28 depletion by qPCR from mice harboring AAV-encoded shRNAs against Trim28 (shTrim28) compared to control, shRNAs against Luciferase (shLuci). (C) Histological examination at the level of the cortex (left panel) and hippocampus (right panel) of mice expressing shTrim28 or shLuci. Cortical and CA1 thickness are measured via cresyl violet staining and astrogliosis is measured using GFAP staining. In (B) and (C), n = 4–6. *** and ns denote p<0.001 and p>0.05, respectively.

Figure 2—figure supplement 2. α-Syn and tau levels are reduced in multiple brain regions from Trim28 adult knockout mice.

Figure 2—figure supplement 2.

(A) Western blot analysis of α-Syn, tau and Trim28 levels in the midbrain from adult knockout mice and littermate controls. (B) Correlation analysis between normalized Trim28 levels and α-Syn or tau levels in hippocampal or midbrain extracts from Trim28 adult knockout mice and littermate controls (obtained from Western blot analysis in Figure 2D and Figure 2—figure supplement 2A). R2 values are presented for each linear regression. (C) qPCR analysis of Trim28 expression from peripheral organs in adult knockout mice and littermate controls. In (A and B), n = 12–13 per group. In (C), n = 7–8 per group. *, ** and **** denote p<0.05, p<0.01 and p<0.001, respectively.