Generation of KCTD10^−/− mice.

<p>(A), the genomic DNA of <i>Kctd10</i> gene includes 7 exons and 6 introns. The genetic manipulation of <i>Kctd10</i> was designed to delete exon 2, the first loxP was inserted into intron 2, and the second loxP together with neomycin-resistant gene flanked by FRT sites was inserted into intron 3. (B–D), Gene disruption was confirmed using Southern blotting in positive ES cells. (E), Genotyping strategy of the mutant mouse, there is only a 670 bp band in KCTD10^−/− mice, two bands (670 bp and 306 bp) in the KCTD10^+/− mice, and only a 306 bp in the wild type mice. (F–G), RT-PCR and western blotting results indicate that the deletion is successful. (F), embryos with the same genotype were collected, and total RNA was extracted separately, reverse transcripted into cDNA and real-time PCR was performed to determine the KCTD10 mRNA levels. (G), embryos with the same genotype were collected, and total protein was extracted separately and immunoblotted by anti-KCTD10 antibody.