Nucleospin genomic dna purification kit

Mice homozygous for the loxP-flanked (floxed) Stat3 gene (STAT3^fl/fl) were kindly gifted from Dr S Akira. Mice carrying a Cre transgene under the control of the distal LysM promoter (LysM-Cre^+/+) were purchased from the Jackson Laboratory (Bar Harbor, ME). Mice with a STAT3 deletion in myeloid cells were generated by crossing mice with the floxed STAT3 allele with mice expressing Cre under the control of the LysM promoter. Genomic DNA was isolated from tail tips using a NucleoSpin genomic DNA purification kit (Macherey-Nagel GmbH, Duren, North Rhine-Westphalia, Germany). The PCR reaction was performed using AccuPower PCR premix (Bioneer, Daejeon, Korea) with the primers, which are specific for exons 22 and 23 of STAT3 and Cre transgene, according to the manufacturer’s instructions. All experiments were performed with male mice aged 8–10 weeks. Experimental animals were maintained under specific pathogen-free conditions and 22±1 °C with a reversed 12 h light–dark cycle (lights on at 0700 h). All experimental procedures were reviewed and approved by the Institutional Animal Care and Use Committee at the College of Medicine, Seoul National University.