Figure 4.

LMIR8 is highly expressed in pDCs. (a) Ba/F3 cells transduced with Flag-tagged LMIR1, LMIR2, LMIR3, LMIR4, LMIR5, LMIR7, LMIR8, or mock were stained with mouse anti-Flag Ab or a control Ab followed by PE-conjugated anti-mouse IgG goat F(ab’)₂ Ab (upper panel) or with biotinylated anti-LMIR8 Ab or biotinylated rat IgG2a Ab followed by PE-conjugated streptavidin. (b–h) Single cell suspensions were prepared from BM (b–f), spleen (g), or lymph node (h). (b–d) Cells were stained with biotinylated anti-LMIR8 Ab or biotin rat IgG2a Ab followed by PE-conjugated streptavidin and FITC-conjugated Abs as indicated. FSC^lowSSC^low populations (b), FSC^highSSC^high populations (c), or FSC^intSSC^int populations (d) were gated. (e–h), BM cells (e,f), spleen cells (g), or lymph node cells (h) were stained with FITC-conjugated anti-CD11c Ab and APC-conjugated anti-B220 Ab (e) or with FITC-conjugated anti-BST2 Ab and APC-conjugated anti-Siglec-H Ab (f–h), and then with biotinylated anti-LMIR8 Ab or biotinylated rat IgG2a Ab followed by PE-conjugated streptavidin. CD11c⁺B220⁺ (e) or BST2⁺Siglec-H⁺ (f–h) cell populations were gated and analyzed for LMIR8 expression. (i) Flt3 ligand-induced BMpDCs, BMmDCs, BMMΦ, or BMMCs were stained with biotinylated anti-LMIR8 Ab or biotinylated rat IgG2a Ab followed by PE-conjugated streptavidin. One representative of four independent experiments is shown.