Molecular cloning of HA⁺ B lymphocytes.

<p>PBMCs from 4 anonymous blood bank donors were stained as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070620#pone-0070620-g004" target="_blank">Figure 4B</a>. Single H1⁺, H3⁺, or H1^negH3^neg CD20⁺ B-cells were sorted to perform molecular cloning and analysis of their paired V_HV_L Ig regions as described in Material and Methods section. <b>A–E.</b> Distribution of V_H (A), D_H (B), J_H (C), V_k (D) and J_k (E) gene use across arrays of B-cells sorted from each donor (16 and 18 HA⁺ clones from donors #1 and #2; 35 HA⁺ and 20 HA^neg clones from donor 3; 16 HA⁺ and 16 HA^neg clones from donor #4. <b>F–I.</b> Number of mutations in H1⁺ and H1^neg CD20⁺ B-cells from donors #3 and #4, which cause dissimilar (F, H) or similar (G, I) amino acid substitutions in V_H (G,I) and V_L (F,H). NS and ** indicate not significant, or significant (p<0.036) difference between mean numbers of mutations by one-way Wilcoxon non-parametric test.