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Fig. 1. Generation of Kcnma1 CKO mice.
(A) Schematic outlining the generation of Kcnma1 CKO mice using the CRISPR/Cas9 system. (B) The mating system of gene-modified mice. (C and D) The targeted fragment of Osterix-Cre and Kcnma1 was amplified by PCR using genomic DNA templates. (E) DNA sequencing map for Kcnma1f/f and Kcnma1f/+ mice. (F) qRT-PCR analysis of Kcnma1 mRNA levels in bone, muscle, kidney and liver from Osterix-Cre(–); Kcnma1f/f and Osterix-Cre(+); Kcnma1f/f mice. (G) Kcnma1 gene expression was significantly down-regulated in primary osteoblasts from Osterix-Cre(+); Kcnma1f/f mice. The Kcnma1 mRNA levels in all tissues and primary osteoblasts from Osterix-Cre(+); Kcnma1f/f mice were normalized to those in Osterix-Cre(–); Kcnma1f/f mice. Data are presented as mean ± SD (n = 3). ***P < 0.001.
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