Fig 1: tDCs Display Lymphoid Characteristics Associated with pDCs(A) TF expression in sorted mouse splenic DC subsets measured by qPCR (n = 3 in 3 exp). Expression represents ??Cq relative to the internal control gene Rpl13a and cDC2s.(B) Sorted mouse splenic DC subsets analyzed by PCR assay for IgH D-J rearrangement. Actin and IgH germline (GL) are also shown (1 representative of 6 exp).(C) EGFP expression in splenic DC subsets from PTCRA-EGFP mice compared with wild-type (WT) mice. Two mice for each condition shown. Numbers indicate frequency of EGFP+ cells.(D) tSNE map of Lin- events from negatively enriched (anti-CD3 and anti-CD19) mouse splenocytes analyzed by CyTOF. Left: manual annotation; right: colored by protein expression. Bottom panels show splenic pre-DCs gated as CD3- CD19- CD335- B220- and CD11c+ MHC class II- CD135+ CD172a- as described (Liu et al., 2009). Pre-DCs were overlaid in the tSNE map (pink dots). See Figure S5 for a comparison between spleen tDCs and BM pre-DCs.(E) Total numbers of tDCs and pre-DCs (n = 5 in 3 exp). The frequency of pre-DCs that overlap tDCs (white) or other cells (black) is shown.(F) Frequency of tDCs that correspond to pre-DCs (average of n = 5 in 3 exp).(G) Surface marker expression in spleen pre-DCs and tDCs representative of 3 exp. Numbers indicate frequency of positive cells.Bar graphs indicate mean ± SD.