Figure 3

Flow cytometric analysis of immune cells trafficking prions from the PC to MedLN 2 HPI.

PBS (rows a–g, v-ab, ac-ai and ax-bd), fluorescent beads (h–n, v-ab, aj-ap and ax-bd) or prion rods (o–u, v-ab, aq-aw, ax-bd) were injected into the PC of mice and cells harvested from peritoneal lavage fluid (a-ab) or mediastinal lymph nodes (ac-bd) two hours later. Graphs in the first column show cells from mice treated with PBS (panels a, v ac and ax), fluorescent beads (h, v, aj and ax) and prion rods (panels o, v, aq and ax). Fluorescent cells (red or green dots) and total cells (grey dots) are plotted to show relative size (forward scatter, linear scale), granularity (side scatter, log scale) and proportion of total live cells that fluoresce. Cells were also stained with antibodies against immune cell surface markers and gated for SSC^hiLy6G- Ly6C+CD11b+ monocytes (panels b, i, p, w, ad, ak, ar and ay), SSC^hiLy6G+ Ly6C-CD11c- CD11b+ neutrophils (c, j, q, x, ae, al, as and az), SSC^hiLy6G- Ly6C-CD11b+CD11c+ DCs (d, k, r, y, af, am, at, ba), SSC^hiLy6G-Ly6C-CD11c-CD11b+ MΦs, (e, l, s, z, ag, an, au and b), SSC^loB220+CD21+CD3- B cells (f, m, t, aa, ah, ao, av and bc) and SSC^loCD21-B220- CD3+ T cells (g, n, u, ab, ai, ap, aw and bd). Cell counts in graphs are shown as log₁₀ per 10⁵ total cells. Horizontal bars below the PC and MedLN panels represent relative proportions of prion-bearing monocytes (red stripe), neutrophils (solid red), DCs (dotted), MΦs (checkered), B cells (white) and T cells (black).