Extended Data Fig. 2: Unc93b1^PKP does not alter TLR9 responses, unlike Unc93b1^D34A.

(a) Representative flow cytometry analysis showing percent TNFα positive cells, measured by intracellular cytokine staining, of indicated RAW macrophage lines after stimulation with CpG-B (25 nM), R848 (10 ng/ml), ssRNA40 (1 µg/ml), PolyIC (20 µg/ml), or LPS (10 ng/ml). Shaded histograms show unstimulated controls. (b) TNFα production, measured by ELISA, from the indicated RAW macrophage lines after stimulation for 8h with R848 (10 ng/ml), CpG-B (25 nM), or LPS (50 ng/ml). Bars show mean of n=4 biological replicates pooled from two independent experiments (unpaired two-tailed Student’s t-test). (c) TLR7 and TLR9 trafficking are normal in Unc93b1^PKP but not in Unc93b1^D34A RAW lines. Immunoblot of TLR7-HA and TLR9-HA from lysates of indicated RAW macrophage lines; FL: full-length. All data are representative of three independent experiments.