Extended Data Fig. 2: Crispr-Cas9 Gabarapl1-KO mouse validation. | Nature Cardiovascular Research

Extended Data Fig. 2: Crispr-Cas9 Gabarapl1-KO mouse validation.

From: Targeted glycophagy ATG8 therapy reverses diabetic heart disease in mice and in human engineered cardiac tissues

Extended Data Fig. 2

a, Gabarapl1 sequence details for the 8 founder Crispr-Cas9 mouse lines confirming excision of exons 2-4 from the Gabarapl1 gene (Next Generation sequencing). Text in red highlights mutations differing from the predicted sequence. b, Validation of Gabarapl1 knockdown using DNA electrophoresis to identify the presence of the Gabarapl1-KO allele in the heterozygote Gabarapl1-KO (tail sample). c, Immunoblot to confirm the absence of the GABARAPL1 band in the homozygote knockout mouse (membrane-enriched fraction of heart homogenate). d, Heterozygote Gabarapl1-KO mice exhibit ~50% knockdown of the Gabarapl1 gene (qPCR) in the heart, and absence of the Gabarapl1 gene is confirmed in the homozygote Gabarapl1-KO mouse heart (male mice, 30 weeks old, WT n = 4, KO/WT n = 5, KO/KO n = 5 animals/group, analyzed by 1-way ANOVA with Bonferroni post-hoc, p < 0.0001). Data presented as mean ± s.e.m. *p < 0.05. Related to Fig. 3.

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