Pristane

MRL/lpr mice were a kind gift from Dr. Yang, Institute of Immunology, Army Medical University, Chongqing, China, and C57BL/6 mice were purchased from The Vital River (Beijing, China). Husbandry environmental conditions consisted of 12‐hours light, 12‐hours darkness, and ambient temperature of 20°C and relative humidity of 30%‐60% at the Third Military Medical University (Chongqing, China). The animal experiments were approved by the Animal Ethics Committee of the Third Military Medical University and were performed according to the international and Chinese guidelines of animal research.
For pristane‐induced SLE model, female C57BL/6 mice, at 10‐12 weeks of age, were given a single intraperitoneal injection of 0.5 mL of pristane (Sigma‐Aldrich) to induce SLE‐like autoimmunity. At the end of the indicated experimental period, animals were killed and specimens including blood, spleen, thymus and kidney were collected.

For the pristane-induced lupus mouse model, 8-weeks old female mice were i.p. injected with 500 μl pristane(Sigma, catalog P9622) and fed with ND or LID for 6 months. After 6 months of pristane stimulation, mice were sacrificed for analysis. The urine protein was detected by a colorimetric assay strip (URIT). The spleen and dLNs (isolated from the inguinal lymph nodes) were collected for flow cytometric analysis, and the renal tissue was fixed in formalin and embedded in paraffin for histological analysis. Serum was collected for auto-antibody analysis.

Pristane-induced lupus-like disease was initiated by injecting 500 µL of 2,6,10,14-tetramethyl-pentadecane or Pristane (Sigma) into the peritoneal cavity of 7–10 weeks-old female mice. For control individuals, genotype- and age-matched female mice were injected with 500 µL of phosphate buffer saline pH 7.4 (PBS) (Gibco). Mice were attributed to PBS- or Pristane-injected groups randomly and maintained in the same cage for the whole procedure. Pristane- and PBS-injected mice were analyzed 8 weeks or 24 weeks after injection.

C57BL/6J WT mice were purchased from Charles River Laboratories (L’Arbresle, France). Lyn^−/−, Mcpt8^{DTR 43}, and Lyn^−/−Mcpt8^DTR mice on a pure C57BL/6 background were bred in our animal facility. For lupus-like disease studies, mice were aged for a minimum of 30 weeks before treatment and analysis (“aged”). For other ex vivo or in vivo analysis, young mice were 8−12 weeks old, unless otherwise specified (“young”). Pristane-induced lupus mouse model was realized by injecting 500 µL of Pristane (Sigma-Aldrich) intraperitoneally (i.p.) or PBS as a control to 8 weeks old C57BL/6J WT mice. These mice were analyzed 24 weeks after injection. Mice were maintained in specific pathogen-free conditions, used in accordance with French and European guidelines and approved by local ethical committee and by the Department of Research of the French government under the animal study proposal number 02484.01.

C57BL/6J wild-type mice were purchased from Charles River Laboratories (Ecully, France). The Rosa26-loxP-Stop-loxP-DTA C57BL/6J $\left(R_{26}^{DTA/DTA}\text{\hspace{0.17em}or\hspace{0.17em}}{R}_{26}^{DTA/+}\right)$ mice (9 (link)) were purchased from The Jackson Laboratory through Charles River Laboratories. Il4^fl/fl C57BL/6J mice were previously described (11 (link)). For all experiments, WT, Mcpt8^CT/CT, Mcpt8^CT/+, Mcpt8^CT/+ x Il4^fl/fl, Il4^fl/fl, $R_{26}^{DTA/DTA}$ , and $Mcpt{8}^{CT/+}\times R_{26}^{DTA/+}$ mice were used on a C57BL/6J, C57BL/6N, or C57BL/6JxN mixed background.
For phenotyping experiments, mice from both sexes in equal proportions were used between 8 and 15 weeks of age. For pristane-induced lupus-like (PIL) disease experiments, 8-week-old female mice of the indicated genotype received a single intraperitoneal injection of 500 µl of pristane (Sigma-Aldrich, Burlington, MA, USA) or phosphate-buffered saline (PBS, Gibco) as a control and were sacrificed 8 weeks later. Mice were maintained under specific pathogen-free conditions in our animal facilities. The study was conducted in accordance with the French and European guidelines and approved by the local ethics committee, comité d’éthique Paris Nord N°121 and the Ministère de l’enseignement supérieur, de la recherche et de l’innovation under the authorization number APAFIS#14115.