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. 2020 Jan 21;2(3):166–181. doi: 10.1096/fba.2019-00091

Figure 1.

Figure 1

Generation, genotyping, and characterization of gas7‐knockout mice. A, The structure of the Gas7 wild‐type and deletion (knockout) allele. For Gas7 gene targeting, we excised the region from exons 5 to 11 of Gas7 genomic DNA and generated a framework error and stop codon in exon 12 that prevented the alternative splicing responsible for forming truncated Gas7. The Gas7 wild‐type and deletion allele were verified by two primer pairs, P1‐P2 and P3‐P4. The predicted PCR product length of the P1‐P2 primers is 492 base pairs (bp) and for the P3‐P4 pair it is 625 bp. White arrows indicate the P1 and P2 primers; black arrowheads indicate the P3 and P4 primers. B, Genotype analysis by PCR was performed on mouse tail genomic DNA. PCR products amplified a ~492 bp fragment in wild‐type and a ~625 bp fragment in knockout mice, and both fragments were found in the heterozygotes. M indicates a 1 kb DNA ladder (Fermentas). C, Western blot analysis. Total brain lysates from wild‐type (WT), gas7 heterozygous (HT), and gas7‐knockout (KO) mice were electrophoresed by 10% SDS‐PAGE. After blotting, the membranes were probed with Gas7 antibody to show that Gas7 protein exists in WT and gas7 HT but not in gas7 KO mice