Nod mice

C57BL/6 (B6) and NOD mice were purchased from Taconic (Ejby, Denmark). B6.E2-2^fl/fl-CD11c.cre [23 (link)] were backcrossed to NOD genetic background for >10 generations to generate NOD.E2-2^fl/fl-CD11c.cre. SNP-based scans were used to ensure that all chromosomes, except chromosome 18 carrying the E2-2 locus were of NOD origin. By in depth SNP analysis of chromosome 18 we have limited the region flanking the E2-2 locus to <19 cM corresponding to <24 Mb (S1 Table). All animals were bred and maintained in a specific pathogen-free environment at the animal facilities at Umeå University or Lund University and all strains were maintained by continuous inbreeding. Mice were sacrificed by cervical dislocation. Only female mice were used for experiments.

All experiments described herein were performed in accordance with the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research and were approved by the Tufts Medical Center Animal Care and Use Committee, the University of Southern California Institutional Animal Care and Use Committee, the Scripps Research Institute Animal Care and Use Committee, and Regierung von Unterfranken (Friedrich Alexander University Erlangen-Nürnberg). MRL/MpJ-Fas /J (MRL/lpr; female, 12 weeks old) and NOR/LtJ (NOD; male, 13 weeks old) mice and their respective age- and sex-matched MRL/MpJ (MRL +/+) and BALB/c control mice were purchased from the Jackson Laboratories (Bar Harbor, ME). We also used in-house bred, 12–13-week-old male NOD mice that were purchased from Taconic Biosciences (Hudson, NY). Animals were euthanized and the exorbital lacrimal glands were harvested and processed for paraffin embedding, total protein extraction, RNA extraction or stimulation with oxytocin. In other experiments, SMA-GFP mice (kind gift of Dr. Ivo Kalajzic), in which lacrimal gland MECs are labeled with GFP, were used to isolate MECs from the lacrimal gland by fluorescence-activated cell sorting (FACS).

C57BL/6, BALB/c, and NOD mice were purchased from Taconic (Hudson, NY, USA). NOD-BDC2.5 and NOD.SCID mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). 129SV-C57BL/6 C/EBPβ^−/− mice were obtained from Dr. Claude Asselin (Université de Sherbrooke) with the agreement of Dr. Peter F Johnson (National Cancer Institute, Frederick, MD, USA). All mice were housed under pathogen-free conditions, in accordance with the guidelines of the Institutional Animal Care Committee of the University of Sherbrooke (Protocols 93-14 and 208-09).

Specific pathogen-free (SPF) NOD mice initially obtained from Taconic Biosciences (Hudson, NY) were bred in the Coverdell animal facility at University of Georgia (UGA). We chose NOD mice, a murine model that spontaneously develops T1D, since they have genetics and changes in immunity that lead to the destruction of pancreatic β-cells similar to humans and are an excellent model for assessing possible environmental factors for T1D risk [14 (link)]. Mice were kept in polysulfone cages with irradiated laboratory animal bedding and Bed-r’Nest for enrichment (The Andersons Inc., Maumee, Ohio) at 22–25 °C with relative humidity 50 ± 20 and a 12-h light/dark cycle. Negligible amounts of BPA have been reported to leach from new or used polysulfone cages maintained at room temperature [15 (link),16 (link)]. Water and food were provided ad libitum. Mice had access to water through the animal facility’s automatic watering system. Animal caretakers were blinded to the exposure groups. In all experiments performed, animals were treated humanely with regard to alleviating animal suffering. An approved animal protocol (A2017 09-001-Y2-A2; Approved on: 2017-09-28) by the UGA Institutional Animal Care and Use Committee (IACUC) was followed for all procedures.