Figure 2.

Unc93 homolog B1 (Unc93b1) controls the cleavage of toll-like receptor 9 (TLR9) in primary immune cells. (A–C) Gating strategy of splenocytes. Splenocytes were stained with indicated antibodies and analyzed by flow cytometry. Gated cells with arrows were drilled down and analyzed by other markers. Whole cDCs were gated out and the other cells were shown in (C). (D) Intracellular TLR9 in B cell, CD8α⁺ cDC, CD8α⁻ cDC, monocytes, and neutrophil were stained by anti-TLR9 (J15A7, open histogram) or IgG1 isotype control (tinted histogram). The surface markers of these cells were shown in (A–C). (E) Form of full length (Full) or cleaved C-terminal of endogenous TLR9. The endogenous TLR9 of bone marrow-derived cDCs from WT, Ifnar1^−/−, Unc93b1^D34A/D34A, Unc93b1^−/−, or Tlr9^−/− mice were immunoprecipitated and detected. β-actin in whole cell lysate (WCL) was detected as an internal control. The intensity of the bands was quantified and shown in the table. At least each four mice (A–D), sex, and age matched were used. At least three times of experiments were performed independently (E). Abbreviations: ND, not detected; N/A, not applicable.