Expression of nAChRs in CTLs from human and mice.

<p>Purified RNA from human and mouse CD8 T cells were examined for nAChR subunits using PCR. A. PCR results (40 cycles, all PCR products were confirmed by sequencing). Memory OT-I cells were obtained as described in the text and as previously reported <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068183#pone.0068183-Xiao2" target="_blank">[35]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068183#pone.0068183-Basu1" target="_blank">[83]</a>. B–C. Quantitative PCR was performed on RNA from human CD8 T cells (B) or murine naive CD8 T cells (C). Expression of all subunits was relative to β2 (the highest expression) in murine naïve CTLs (C), which was designated as 1. D. Purified naïve OT-I cells were stimulated with 2SI or 3SI (2SI plus IL-12) for 3 days, and expression of nAChRs was compared relatively to β2 in each treatment (2SI and 3SI). E. The expression of 3 dominant nAChR subunits in 2SI and 3SI treatments was compared to naïve OT-I using the housekeeping gene GAPDH as internal control. Subunits α1, α7 and β3 were not detectable in D–E. F. CD8 OT-I cells were purified from naïve OT-I mice or OT-I mice treated with nicotine water at 200 µg/ml for 60 days to quantify the expression of nAChR subunits. Data are representatives of at least three experiments with similar results.