Characterization of MICL Expression in Clec12A Knockout Mice

MICL knockout mice (Clec12A^−/−) on a C57BL/6 background were produced by Taconic Artemis, USA (see online supplementary figure S1A). Clec12A^−/− and wild-type (wt) mice were bred and maintained at the Medical Research Facility, University of Aberdeen. Mice were separately housed in groups and provided freely with food and water. All experimentation conformed to the terms and conditions of UK Home Office licences for research on animals (PPL 60/4007) and the University of Aberdeen ethical review committee.
Characterisation of MICL expression in 8–12-week-old wt and Clec12A^−/− mice was performed by flow cytometry on cells isolated from the peripheral blood, bone marrow, peritoneal cavity, spleen and lungs, as previously described.14 (link) Antibodies used in these experiments included biotin anti-MICL monoclonal antibody 3097 (link) and isotype control, as well as biotin-Gr-1, FITC-7/4, PE-F480, biotin-F480, PerCpCy5.5-CD11b, PE-CCR3, biotin-NK1.1, PE-CD49, PerCpCy5.5-B220, PE-CD19, PerCpCy5.5-CD3, biotin-CD4, FITC-CD8, biotin-CD11c, PerCpCy5.5-Gr-1, APC-Ly6G, PE-CD11b, PE-Ly6G, PECy7-CD11b, Alexa Fluor 488 anti-STAT5, APC-streptavidin (all from BD Biosciences), and Alexa Fluor 700-F4/80 (BioLegend). Flow cytometry was undertaken using a BD LSRFortessa cell analyser and data analysed using FlowJo.