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Fig. 5 | BMC Molecular Biology

Fig. 5

From: A protocol for custom CRISPR Cas9 donor vector construction to truncate genes in mammalian cells using pcDNA3 backbone

Fig. 5

Western blot screening for putative FOXO3 truncation mutants. Total protein lysates were prepared from 50 putative FOXO3 mutant-containing cells and were examined by western blot analyses; employed antibodies are indicated. Wild-type FOXO3 was approximately 80 kDa, whereas, mutant FOXO3 protein was approximately 45 kDa. This second proof of principle screen examined 50 potential FOXO3 mutant clones. Three of these were homozygous mutant (16, 25 and 43), three appeared to be heterozygous (14, 30 and 31) and 44 were wild-type. The homozygous mutants were confirmed by Sanger sequencing

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