Generation of integrin β7 ⁺ PC s through pre- or post-GC response. (A) Experimental design for time-course analysis of generation of GC-dependent PCs in SPL. (B) The frequency of integrin β7^hi in tdTomato⁺IgG1⁺ SPL PCs (left) or the number (right) of tdTomato⁺IgG1⁺integrin β7⁺ SPL PCs (right) at the indicated time points after immunization with NP-CGG (day 9: n = 3, day 10: n = 4, day 11: n = 3, and day 12: n = 2). (C) Experimental design to analyze PCs that are generated through pre-GC (upper) or GC reaction (lower). (D) Gating strategy to analyze IgG1⁺integrin β7⁺SPL or BM PCs that were derived from pre-GC (CD138^hiTACI^hiCD45.1⁺tdTomato⁻) or post-GC response (CD138^hiTACI^hiCD45.1⁺tdTomato⁺) (left) and the frequency of integrin β7^hi in pre-GC PCs (day 7 tdTomato⁻) or post-GC PCs (day 12 tdTomato⁺) in SPL or BM (right, n = 5). (E) Experimental design to analyze generation of integrin β7⁺ PCs in a non-BCR transgenic setting. (F) Representative FCM plots showing integrin β7 expression on CD138^hiTACI^hiNP⁺IgG1⁺tdTomato⁺ cells in SPL or BM. CD138⁺ cells were pre-enriched before surface staining. Data in D and F are representative of two independent experiments. PC, plasma cell; SPL, spleen; BM, bone marrow.