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. Author manuscript; available in PMC: 2018 Nov 21.
Published in final edited form as: Immunity. 2017 Nov 14;47(5):913–927.e6. doi: 10.1016/j.immuni.2017.10.006

Figure 5. Programmed macrophages have a higher activation threshold for TLR7 and TLR9 responses.

Figure 5

(A) Tim-4+ pMacs were stimulated with R848 or ACs for 30min, and phospho-ERK1/2 signal was measured by flow cytometry. Data are representative of three independent experiments. (B) Negative regulators of TLR signaling that are significantly (p < 0.05) reduced in deprogrammed Tim-4+ pMacs relative to programmed Tim-4+ pMacs as measured by RNA sequencing. Results of two independent experiments are shown with replicates averaged. (C–E) TNF production by programmed and deprogrammed Tim-4+ pMacs stimulated with increasing doses of TLR ligands. Data are representative of at least three independent experiments. (D) To compare CpG responses, Tim-4+ pMacs were isolated from RosaTlr9 mice. (F and G) Pleural (F) or lung (G) cells from WT mice were harvested and incubated overnight or for 60h before stimulation. Data are representative of two independent experiments. See also Figure S5.