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. 2013 Dec 1;126(23):5477–5489. doi: 10.1242/jcs.137026

Fig. 1.

Fig. 1.

Generation of the mouse line in which residue I7097 of nebulin was replaced with stop codons in all reading frames, resulting in deletion of the nebulin SH3 domain (NebΔSH3 mice). (A) A restriction map of the relevant genomic region of nebulin (top), targeting construct (middle) and the mutated locus after recombination (lower). Neo, neomycin resistance gene; DTA, diphtheria toxin A chain. Long boxes represent FRT sites and the black box represents the probe used for Southern blot analysis. (B) Detection of WT and targeted alleles by Southern blot analysis on BamHI-digested electroporated ES cells using the probe shown in A. The 8.7 and 6.7 kb bands represent the WT and the targeted allele, respectively. (C) RT-PCR analysis on total RNA isolated from TA muscle using the genotyping primers, confirming the correct targeting of the nebulin gene. (D) Western blot analysis of TA muscle from WT and homozygous NebΔSH3 mice using antibodies against the nebulin SH3 domain, nebulin M160–164 and nebulin M161–165. α-Actinin antibody was used as loading control. (E) Western blot analysis of heart (H) and TA muscle from WT and homozygous NebΔSH3 mice using antibodies against nebulette. α-Tubulin antibody was used as loading control. M, mutant allele.