Extended Data Fig. 1: CD40 stimulation induces CD70 and 4-1BBL expression in cDCs.
From: Mechanisms of CD40-dependent cDC1 licensing beyond costimulation
a, Hierarchical clustering of 89 genes induced at least two-fold in SDLN cDC1s treated in vitro in the absence or presence of agonistic αCD40 antibody (results averaged from three independent experiments). b, Day 8 Flt3L-treated WT (B6) bone marrow cultures were treated with no stimulation, agonistic αCD40 antibody, poly(I:C), or both. cDC1 surface expression of CD40 (top), CD70 (middle) and 4-1BBL (bottom) was analyzed by flow cytometry after 24 h. Pre-gate: B220− Siglec H− MHC class II+ CD11c+ XCR1+ Sirpα-. Numbers represent the percentage of cells in the indicated gates. Data represent two independent experiments. c, d, f, WT mice were injected i.p. with PBS, agonistic αCD40 antibody, poly(I:C), or both. c, Spleens were harvested 24 h after injection and cDC1 were analyzed for surface expression of CD40 and CD70 by flow cytometry. Pre-gate: B220− F4/80− MHC class II+ CD11c+ XCR1+ Sirpα-. Numbers represent the percentage of cells in the indicated gates. Data represent two independent experiments. d, SDLN were harvested 24 h after injection and cDC1 were analyzed for surface expression of CD40 and CD70 by flow cytometry. Pre-gate: B220− CD326− MHC class II+ CD11cint XCR1+ Sirpα-. Numbers represent the percentage of cells in the indicated gates. Data represent two independent experiments. e, Day 8 Flt3L-treated bone marrow cultures were treated with no stimulation, agonistic αCD40 antibody, poly(I:C), or both. cDC2 surface expression of CD40 and CD70 were analyzed by flow cytometry after 24 h. Pre-gate: B220− Siglec H− MHC class II+ CD11c+ XCR1−Sirpα +. Numbers represent the percentage of cells in the indicated gates. Data represent two independent experiments. f, SDLN were harvested 24 h after injection and cDC2 were analyzed for surface expression of CD40 and CD70 by flow cytometry. Pre-gate: B220− CD326− MHC class IIhi CD11cint XCR1− Sirpα +. Numbers represent the percentage of cells in the indicated gates. Data represent two independent experiments.
