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. Author manuscript; available in PMC: 2020 Mar 23.
Published in final edited form as: Nature. 2019 Sep 23;575(7782):371–374. doi: 10.1038/s41586-019-1611-7

Extended Data Fig. 8: Tethering of TLR9 and Unc93b1.

Extended Data Fig. 8:

(a) Cysteine mutants of TLR9 and Unc93b1 do not affect trafficking of TLR9 to endosomes. Immunoblot of TLR9-HA from macrophage lines expressing the indicated TLR9-HA and Unc93b1-Flag cysteine mutants. Representative of two independent experiments. (b) TLR9-HA immunoblot under non-reducing conditions after immunoprecipitation of TLR9-HA from macrophage lines shown in (a). The high molecular weight band indicates disulfide-bond formation between Unc93b1 and TLR9. Representative of two independent experiments. (c) Unc93b1-tethered TLR9 is unable to signal. NF-κB luciferase assay in HEK293T cells expressing the indicated cysteine mutant combinations and stimulated with CpG-B (1µM) for 16h. Data are normalized to Renilla expression and expressed as luciferase fold change over unstimulated controls. Data are mean ± s.d., n=3 biological replicates. p-values determined by unpaired two-tailed Student’s t-test. Representative of three independent experiments. FL: full-length.