<p>(A, B) CD43<sup>-</sup> splenic B cells from Bim<sup>-/-</sup> mice were treated with anti-IgM F(ab’)<sub>2</sub> (15μg/ml) for various times in the presence or absence of Syk kinase inhibitor R406 (4μM), Bruton’s tyrosine kinase inhibitor Ibrutinib (PCI-32765) (20nM), or TORC1 inhibitor rapamycin (50nM). Whole cell extracts were fractionated by SDS-PAGE and Mcl-1 protein was analyzed by immunoblotting. β-actin was used as a loading control to normalize between samples. (C, D) CD43<sup>-</sup> splenic B cells from BL6, BCAP<sup>-/-</sup> (C) or CD19<sup>-/-</sup> (D) mice were cultured for the indicated times with or without anti-IgM F(ab’)<sub>2</sub> (15μg/ml). Whole cell extracts were prepared and assayed for Mcl-1 levels by Western blot analysis. β-actin was used as a loading control to normalize between samples. Representative gels from 3 independent experiments are shown for A and B and 2 independent experiments for C and D. Cell viability profile of BCAP<sup>-/-</sup> and CD19<sup>-/-</sup> B cells are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0146955#pone.0146955.s004" target="_blank">S4 Fig</a>.</p
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