Staining with various cell surface antigens and 2E6 on freshly isolated splenic lymphocytes. (A) Staining of freshly isolated splenocytes with 2E6 mAb. Freshly isolated splenocytes from BALB/c mice were stained with various Abs (CD11c, B220, Ly6C, CD45RB, Gr-1, I-Ad, DX5, CD19, CD14, CD11b, and CD3) in combination with 2E6 mAb. (B) Morphologic features of 2E6⁺ cells. CD3⁺, CD19⁺, CD11b⁺, CD14⁺, and DX5⁺ cells were depleted from freshly isolated spleen cells by Auto MACS. The 2E6⁺ cells were then sorted by cell sorter (BD Biosciences). Cells were plated onto slides by centrifugation and stained with May-Giemsa (original magnification, × 1000). Micrography was performed using a Nikon OPTIPHOT-2 microscope equipped with a Plan 40 objective lens, the numerical aperture of which was 0.70 (Nikon, Tokyo, Japan). Images were acquired using a COOLPIX 990 3.34-megapixel camera (Nikon) and Preview software 2.1.0 (Apple Computer, Cupertino, CA).