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. 2017 Jul 17;114(31):E6371–E6380. doi: 10.1073/pnas.1706504114

Fig. 1.

Fig. 1.

DC subpopulations in Vps34f/f;CD11c-Cre mice. (A) Spleens and a pair of inguinal lymph nodes from representative 12-wk-old Vps34f/f and Vps34f/f;CD11c-Cre mice are shown. (Scale bar, 1 cm.) (B) Single-cell suspensions of spleens and a pair of inguinal lymph nodes were prepared and counted. Results from three independent experiments (five to seven animals per group) are shown. *P < 0.001. (C) Percent and absolute numbers of DCs in the spleen and lymph nodes of Vps34f/f or Vps34f/f;CD11c-Cre mice. Results from three independent experiments (five to seven animals per group) are shown. *P < 0.01. (D) Splenocytes from 12-wk-old Vps34f/f or Vps34f/f;CD11c-Cre mice were stained with anti-CD11c, -CD103, -CD8α, and -CD11b antibodies. The CD11chiCD11blo cells were evaluated for CD103 expression, and levels of CD8α on CD103− and CD103+ cells were measured. (E) Summary of the percentage and absolute numbers of CD103- or CD8α-expressing DCs in the spleen. Pooled results from two independent experiments (five to seven mice in each group) are shown. *P < 0.01. (F and G) Single-cell suspensions of lungs from the indicated mice were stained with anti-CD11c, -CD103, -CD8α, and -CD11b antibodies. A representative experiment (F) and a summary of the data pooled from two independent experiments (six mice in each group) (G) are shown. (H) Frequency of CD8α+ DCs in 4-wk-old mice. (I) Bone marrow chimeras were generated in lethally irradiated B6 mice (CD45.2) by transfer of 107 cells bone marrow cells from wild-type B6.SJL mice (CD45.1) and Vps34f/f;CD11c-Cre (CD45.2) or Vps34f/f (CD45.2) mice mixed at a 1:1 ratio. At the end of 12 wk spleen cells from the chimeric mice were tested for the frequency of CD8α+ cells among CD45.2+ DCs. A summary of two independent experiments (five mice per group) is shown. *P < 0.05. (J) Bone marrow-derived Flt3L-driven DCs were generated in the presence of 150 ng/mL of rFlt3L for 9–21 d of in vitro culture. Cells analogous to splenic CD8α+ DCs were identified as CD45RA−CD24+Sirp-α− cells. Representative plots from three individual experiments with two mice per group are shown.