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. 2020 Jan 6;9(1):133. doi: 10.3390/cells9010133

Figure 2.

Figure 2

Generation of conditional TCTP knockout mouse model. (A) Diagram of the targeting strategy for the generation of the conditional TCTP allele. The targeting construct contains loxP sites (arrowheads) flanking exons 3–4 and restriction sites EcoRI (E) and HindIII (H). (B) Genotyping with PCR, top panel, and Western blotting of TCTP expression, bottom panel, from Nestin+/+; TCTPflox/flox (Control), NestinCre/+; TCTPflox/flox (heterozygous), and NestinCre/+; TCTPflox/+ (cKO) mice at P0.5. (C) qRT-PCR of relative TCTP mRNA expression in the whole brain from control, heterozygous, and conditional knockout mice at different stages was estimated (data presented as mean ± SEM, * p < 0.05, *** p < 0.001 compared with control, # < 0.05 compared with E13.5 cKO, one-way ANOVA, n = 3–11 per group). (D) Immunohistochemistry of TCTP protein from control and TCTP-cKO mice at P0.5. DG, dentate gyrus; Hipp, hippocampus; SVZ, subventricular zone. Scale bars: a, e, 400 μm; b, f, 200 μm; c, g, 40 μm; d, h, 100 μm. (E) Sections at E16.5 were also detected by double-labeled immunofluorescence with antibodies against TCTP to identify the deletion and Nestin to identify neural precursor cells. A representative image is shown for three independent experiments. Scale bars: 100 μm for right panel, 200 μm for left panel.