Figure 1.

Expression patterns of Vgll2 mRNA expression in several muscles, and generation of Vgll2-deficient mice. (a) Vgll2 mRNA levels were measured by qPCR in the soleus (SOL), gastrocnemius (GAS), and extensor digitorum longus (EDL) muscles from 12-week-old Vgll2 ^+/+ and Vgll2 ^−/− mice (n = 6). *P < 0.05 vs. Vgll2 ^+/+ soleus muscles. Data are presented as mean ± SEM. For comparison, the expression level in Vgll2 ^+/+ soleus was arbitrarily set at 1. (b) Schematic diagram showing the strategy used to generate the Vgll2 null allele. The top row depicts the wild-type Vgll2 allele, which consists of three exons (solid boxes). The initiation Met codon for Vgll2 exists in exon 1. The second row depicts the targeting construct. The third row depicts the mutated Vgll2 allele. The lacZ expression cassette with a SV40 polyadenylation signal, which was followed by the loxP-flanked puromycin resistant gene expression cassette (Puro) in the reverse orientation, was fused to the initiation codon of Vgll2. The diphtheria toxin A expression cassette (DT-A) was included at the 5′-end of the targeting vector. The external probe used for Southern blot analysis lies outside the 5′-homologous arm (gray box). RI, EcoRI; RV, EcoRV. (c) Southern blot analysis for genotyping wild-type (Vgll2 ^+/+), heterozygous Vgll2-deficient (Vgll2 ^+/−), and homozygous Vgll2-deficient (Vgll2 ^−/−) animals. Digestion with EcoRI of Vgll2 ^+/+ and Vgll2 ^−/− alleles gave rise to fragments of 2.2 and 4.6 kb, respectively. An uncropped original image is shown in Supplementary information. (d) Vgll2 protein levels were assessed by Western blotting using nuclear protein extracts obtained from the soleus muscle of 12-week-old Vgll2 ^+/+ and Vgll2 ^−/− mice. Histone H2B served as a loading control. Analyses were performed on four mice per genotype. Data from two mice per genotype are shown. Uncropped original images are shown in Supplementary information.