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. 2011 Jun 3;6(6):e20412. doi: 10.1371/journal.pone.0020412

Figure 5. Tacrolimus decreased SFRP2 induced nuclear NFATc3 in 2H11 endothelial cells.

Figure 5

A) 2H11 cells were treated with control (1.5% DMSO), mouse recombinant SFRP2 (7 nM)+1.5% DMSO; or SFRP2 7(nM)+tacrolimus 10 µM in 1.5% DMSO for 1 hour, and nuclear protein lysates were collected and analyzed by Western blot analyses probing for NFATc3 as described in “Material and Methods”. The loading control was TATA binding protein TBP antibodies (a nuclear marker). SFRP2 increased nuclear NFATc3 compared to control cells (p = 0.003). Full-length blots/gels are presented in Figure S2C. B) The experiment was repeated as above except that whole cell lysates rather than nuclear lysates were extracted and analyzed by Western blot analyses probing for NFATc3. The loading control was beta-tubulin. Tacrolimus did not inhibit total NFATc3 protein. Taken together this shows that tacrolimus inhibits SFRP2 induced NFATc3 nuclear translocation but not total protein levels.