Fig 1: Ppef2 is predominantly expressed by CD8+ cDC1. (A) Gene expression profiling of spleen and blood cells after sorting. Shown is the quantitative real-time PCR result for Ppef2 of three independent sort experiments (n = 3 pooled mice per sort) ± SEM analyzed by the ?Ct method with HPRT as housekeeping gene. Cells were identified and sorted as ESAMhi (CD11c+MHCII+CD11b+ESAMhi); ESAMlo (CD11c+MHCII+CD11b+ESAMlo); CD8+DC (CD11c+MHCII+CD11b-CD8+); pDC (CD11b-SiglecH+); CD4+ T cells (CD3+CD4+); CD8+ T cells (CD3+CD8+); B cells (CD19+B220+); blood monocytes NK1.1-B200-CD115+CD11b+Ly6G- cells with differential Ly6C and MHCII expression as indicated. Gating strategies are shown in Supplemental Figure 1B. (B) Ppef2 expression in GM-CSF cultured BMDCs of C57BL/6 mice after cell sorting on day 7 of culture based on the expression of CD11c, MHCII, and CD86. Error bars represent SEM of 3 independent experiments. (C) Ppef2 expression 16h after in vitro stimulation with the indicated TLR-ligands of GM-CSF- or Flt3L-cultured BMDCs of C57BL/6 mice, as well as spleen CD8+ DCs 16h after intravenous injection of 10 µg LPS as determined by qPCR. Bar graphs with SEM represent pooled data from independently performed cell cultures [GM-CSF BMDCs: unstim., LPS (n = 5); Flagellin, Poly(I:C), Pam3CSK4, CLO97, (n = 4); anti-CD40, LPS+anti-CD40 (n = 3); Flt3L BMDCs (n = 3); sorted CD8+ DCs (n = 4)]. Statistical analysis was performed using Student's t-test, with *p < 0.05; **p < 0.01; ***p < 0.001.