Animals were used under protocols approved by local institutional research committees and in accordance with UK Home Office guidelines. C57BL/6 (B6) mice were bred in-house. Homozygous CD11c.DTR (B6.FVB-Tg(Itgax-DTR/ EGFP)57Lan/J) mice were bred in-house. Rag2−/− mice were bought from Jackson Laboratory and crossed in-house to CD11c.DTR mice. CD11c.DTR and CD11c.DTR.Rag2−/− syngeneic chimeras were generated as described previously.38 (link) CD11c+ cells were depleted upon intraperitoneal injection of 100 ng of DT (Sigma, UK) in PBS according to published protocols. Chimeras received single injections or repeated injections once every 72 hr, and depletion was assessed in control animals 24–48 hr after injection of DT.
B16.F10 melanoma cells were cultured in RPMI 1,640 medium (Lonza), supplemented with 10% fetal calf serum (FCS; Biosera), 1% L-glutamine (2 mM), and 1% penicillin/streptomycin (100 U/mL). C57BL/6 mice were injected subcutaneously with 1 × 105 B16.F10 cells in 100 μL of PBS. Tumors became palpable by days 5–10. Tumor size was assessed using calipers every 2–3 days. Mice were euthanized if tumors grew to >15 mm in any direction, as per home office regulations.
B16.F10 melanoma cells were cultured in RPMI 1,640 medium (Lonza), supplemented with 10% fetal calf serum (FCS; Biosera), 1% L-glutamine (2 mM), and 1% penicillin/streptomycin (100 U/mL). C57BL/6 mice were injected subcutaneously with 1 × 105 B16.F10 cells in 100 μL of PBS. Tumors became palpable by days 5–10. Tumor size was assessed using calipers every 2–3 days. Mice were euthanized if tumors grew to >15 mm in any direction, as per home office regulations.
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