Fig. 1

Generation of TALEN-mediated CXCL1 knockout mice. A DNA-binding sequences are presented in red or green, and the spacer region for CXCL1-TALEN where a double-strand break will occur is underlined. B T7 endonuclease I (T7EI) assays were conducted using genomic DNA from the founder mice. The arrow shows the size (300 bp) of T7EI-digested DNA fragments. #1, #2, and #3 are the mutant founder (F0) mice generated by injection with CXCL1-TALEN mRNA. C DNA sequences of the CXCL1 locus from live founder mice identified by T7E1 assays in B. “-” shows the deleted nucleotides. D Peripheral blood mononuclear cells and splenocytes were collected from wild-type or CXCL1^−/− mice. Numbers of CD45⁺ and Gr.1⁺ cells were quantified via flow cytometry. E Peripheral blood mononuclear cells were collected from wild-type or CXCL1^−/− mice, CXCR2 expression in neutrophils from WT and CXCR2^−/− mice were analyzed via flow cytometry. F mRNA expression levels of TLR4 and CXCR2 in the brain of WT or CXCL1^−/− mice were analyzed by RT-PCR. G Wild-type and CXCL1^−/− mice were treated with i.p. or i.c.v. LPS injection. Four hours later, levels of TNF-α and IL-6 in the brain tissue or plasma were determined by ELISA. Data are expressed as the mean ± SEM, n = 4 mice in all of the groups