Figure 1.
Figure 1. Generation of mice lacking nesprin 1 CH domains. (A) Schematic of syne1 gene with primer locations used for PCRs in D and E (arrows). (B) Construct used for targeting the syne1 gene, with the exon 9F (yellow rectangle) flanked by two LoxP sites (arrowheads). DTA, diphtheria toxin A; black rectangles, flippase recombination target sites; Neo, neomycin cassette. (C) Southern blot confirmation of the WT allele at 18 kb and presence of the mutant allele (MUT) at 6 kb. (D) Semiquantitative RT-PCR of WT and nesprin 1ΔCH mRNA isolated from skeletal muscle. Note that similarly to WT and as expected, the other nesprin 1 isoforms were present in nesprin 1ΔCH mRNA. (E) qRT-PCR of mRNA from WT and nesprin 1ΔCH using primers specific to the CH domain–encoding exon 9F. Note the significantly decreased levels of nesprin 1ΔCH domain–containing exon 9F compared with WT. **, P < 0.01 according to an unpaired Student’s t test.

Generation of mice lacking nesprin 1 CH domains. (A) Schematic of syne1 gene with primer locations used for PCRs in D and E (arrows). (B) Construct used for targeting the syne1 gene, with the exon 9F (yellow rectangle) flanked by two LoxP sites (arrowheads). DTA, diphtheria toxin A; black rectangles, flippase recombination target sites; Neo, neomycin cassette. (C) Southern blot confirmation of the WT allele at 18 kb and presence of the mutant allele (MUT) at 6 kb. (D) Semiquantitative RT-PCR of WT and nesprin 1ΔCH mRNA isolated from skeletal muscle. Note that similarly to WT and as expected, the other nesprin 1 isoforms were present in nesprin 1ΔCH mRNA. (E) qRT-PCR of mRNA from WT and nesprin 1ΔCH using primers specific to the CH domain–encoding exon 9F. Note the significantly decreased levels of nesprin 1ΔCH domain–containing exon 9F compared with WT. **, P < 0.01 according to an unpaired Student’s t test.

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