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. Author manuscript; available in PMC: 2013 Nov 15.
Published in final edited form as: J Immunol. 2012 Oct 12;189(10):4816–4824. doi: 10.4049/jimmunol.1202165

Figure 4.

Figure 4

Flow cytometry analysis of bNAb-expressing WEHI231 B cells cultured with JRFLΔCT-EGFP pseudovirions. bNAb BCR genes were induced one day before stimulation and then cells were treated for 18 hr as indicated. (A) Analysis of CD69 upregulation in cells stimulated with doxycycline medium alone (control), anti-IgM (MAb M41, 2μg/ml) or 100-fold concentrated JRFLΔCT-EGFP pseudovirus (final concentration 3×109/ml). (B) Cell associated virion binding/internalization as measured by EGFP level. (C) Downregulation of hCκ expression in virion-treated compared to untreated cells. (D–F) Dose/response analysis in b12 and gl-b12 WEHI-231 cell lines after pseudovirion stimulus (highest final concentrations of pseudovirions were approximately 3×109/ml for JRFLΔCT and 2×1012/ml for BaL, corresponding to 1 and 67 nM Env, respectively): (D) Virus binding measured by GFP fluorescence, (E) CD69 induction, (F) Human Igκ downregulation.