Fig. 3.
Effect of 2ME2 on XBP-1 expression.
MM.1S cells were treated with 3 μM 2ME2 for the indicated times. (A) Total cellular RNA was subjected to Northern blot analysis. Data shown is representative of 3 independent experiments with similar results. (B) Total cell lysates were separated by 12.5% SDS-PAGE and analyzed by immunoblotting (IB) with anti–XBP-1 (upper panel) or anti-SHP2 (lower panel) Abs. (C) Purified patient MM cells (CD138+) were treated with 2ME2 for the indicated times and analyzed for mRNA levels by Northern blotting. The blots were reprobed with GAPDH to confirm equal loading. (D) Total cell lysates from purified MM cells (CD138+) (patient 1 and patient 2), normal BM cells, and 2ME2-treated patient cells were separated by 12.5% SDS-PAGE and analyzed by immunoblotting (IB) with anti–XBP-1 (upper panel) or anti-SHP2 (lower panel) Abs.