Fig. 5
HlaH35A induces Notch2-dependent cDC2s via ADAM10-Notch signaling. a Representative flow cytometry histogram (left) and quantification (right) of MHCII+ cell frequencies gated on CD11c+ cells from Fl-BMDCs after 7.5 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 6 per group). The data were pooled from two independent experiments. b Quantification of Ccr7 mRNA expression by qPCR in Fl-BMDCs after 7.5 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 3). c Representative flow cytometry plots (left) and quantification (right) of cDC subset frequencies in Fl-BMDCs after 12 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 6 per group). The data were pooled from two independent experiments. d Quantification of cDC1-associated Toll-like receptor mRNA expression by qPCR in Fl-BMDCs after 6 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 3). e Quantification of cDC2-associated Toll-like receptor mRNA expression by qPCR in Fl-BMDCs after 6 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 3). f Quantification of Irf4 or Klf4 mRNA expression by qPCR in Fl-BMDCs after 6 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 3). g Representative flow cytometry histogram (left) and quantification (right) of ESAM+ cell frequencies gated on CD11c+ MHCII+ SIRPa+ cells in Fl-BMDCs after 12 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 6 per group). The data were pooled from two independent experiments. h Serum levels of IL-23 secreted from Fl-BMDCs after 12 h of treatment with PBS, PA0833, HPF, HPF with GI254023X or HPF with DAPT (n = 6 per group). The data were pooled from two independent experiments. Each data point indicates a biological replicate in (a, c, g, and h). Each data point indicates a biological replicate derived from the average of three technical replicates in (b,d, e, and f). The data are presented as the means ± s.e.m.s. Statistical significance was tested via one-way ANOVA followed by Tukey’s multiple comparisons test (a, b, f, g and h) and two-way ANOVA followed by Tukey’s multiple comparisons test (c–e)
