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. 2018 Aug 1;8:11532. doi: 10.1038/s41598-018-30011-w

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Validation of the Lifr-knockout allele. (A) Schematic of WT and KO alleles detailing the location of the genotyping primers and expected PCR product sizes. (B) Representative PCR analysis of genomic DNA isolated from tail-tip biopsies of neonatal mice identified wild-type (WT), heterozygous (HET) and homozygous (KO) animals. (C) Western blot analysis of neonatal brain tissue homogenates confirmed LIFR protein expression was abolished in KO animals. Tubulin-alpha (TUBA) was used as a loading control. Both LIFR and TUBA bands have been cropped from a single gel blot image which is included in the supplementary materials with the cropped areas highlighted. (D) Transgene inheritance in offspring derived from heterozygous matings based on genomic PCR of DNA isolated from ear-clip biopsies collected at weaning. Chi-squared analysis revealed a significant deviation from the expected Mendelian ratios (X²; p < 0.0001).