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. 2017 May 1;10:121. doi: 10.3389/fnmol.2017.00121

Figure 2.

Figure 2

Generation of Ankfy1/+ mutant mice. (A) Schematic representation of the targeting constructs. The internal ribosome entry site (IRES)-βgeo-polyA cassette was inserted into intron 4 of the ANKFY1 locus. The IRES-βgeo-polyA cassette is presented as a gray box. It contains the fusion gene βgeo(lacZ-neo) placed under the control of an IRES. Arrowheads indicate the primers used for protein coupled receptor (PCR). (B) PCR analyses. Solid arrows indicate bands corresponding to the WT locus, whereas open arrows indicate bands corresponding to the targeted locus. +/+, WT mice; +/−, heterozygous mutant mice. (C) Birth rates of WT mice were approximately half of the expected heterozygous Mendelian ratios (1:2). (D) RT-qPCR analyses of Ankfy1 mRNA levels in the cerebella of WT and Ankfy1/+ mutant mice (*p < 0.05, **p < 0.01). (E) Cerebella from different adult mice were immunoblotted with a polyclonal mouse antibody against Ankfy1. The levels of the β-actin protein in the lysates are shown as an internal control (***p < 0.001). The data represent means ± standard errors of the means (SEM).