The electrophoretic mobility of PRX3 is modified by TS.

<p>A) HM cells were treated with the indicated concentration of TS for 18 hr, and cell extracts were prepared, heated in SDS sample buffer at 95°C for 10 min, resolved by gel electrophoresis, and PRX3 expression was assessed by immunoblotting. Treatment of HM cells with increasing doses of TS caused the formation of a modified species of PRX3 that migrated with an apparent molecular weight of about 45–50 kD. B) Recombinant human PRX3 (rPRX3) was incubated with 10 µM TS in solution in vitro using conditions described by Chiu and colleagues <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039404#pone.0039404-Chiu1" target="_blank">[43]</a>, and samples were resolved by gel electrophoresis and examined by immunoblotting. Pre-treatment of rPRX3 with sodium cyanoborohydride (NaCNB) accentuated the reactivity of TS (lane 2) whereas pre-incubation with NAC inhibited it (lane 5). Addition of TS and N-ethyl-maleimide (NEM) or NAC to rPRX3 treated with NaCNB resulted in multiple species of immunoreactive PRX3 (lanes 3 and 4), whereas treatment with NACNB alone (lane 7) had no effect. C) HM cells were treated with the indicated concentration of TS for 18 hr, and cell extracts were prepared in the absence of reducing agents as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039404#pone.0039404-Phalen1" target="_blank">[69]</a>. Samples (20 µg protein/lane) were resolved under denaturing but not reducing conditions, and the relative distribution of PRX3 monomers to dimers was assessed by immunoblotting.