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. 2014 Sep 23;4:6444. doi: 10.1038/srep06444

Figure 2. OSM induces STAT-3 activation and migration in astroglioma cells.

Figure 2

(A) Whole cell lysates from the CRT-MG, U251-MG and U87-MG cells treated with OSM (10 ng/mL) for 30 min were analyzed by immunoblotting against total STAT-3 and p-Y705-STAT-3. Tubulin was used as the loading control. Cropped blots are used. Full-length blots are shown in supplementary Figure 1. (B) Nuclear and cytoplasmic extracts from CRT-MG cells treated with hOSM (10 ng/ml) for 30 min and analyzed by immunoblotting against total STAT-3 and p-Y705-STAT-3. Tubulin and Histone were used as loading and purity controls of each cellular fraction. Data shown are representative of at least three experiments. P, phospho; CE, cytoplasmic extracts; NE, nuclear extracts. Full-length blots are shown in supplementary Figure 1. (C) Representative micrographs of the scratch wound healing assay in CRT-MG cells in the absence or presence of either OSM or AG490 at 48 h, as indicated. Bar = 100 μm. (D) Confluent CRT-MG cells were assayed for migration into the wound at 48 h after scratching, in the absence or presence of OSM and AG490, as indicated. Migrating cells were measured by counting the cell numbers. The results are expressed as the mean ± SD values (compared with untreated controls at 48 h time point); normalized values are shown. The data shown are representative of three independent experiments. (*P < 0.05 vs. untreated control; †P < 0.05 between OSM-treated cells and OSM/AG490-treated cells.).