Fig. 5.

Knockdown of JAK1 expression or inhibition of JAK1 inhibits TROY-induced GBM cell migration. (A) Migration assay for T98G and T98G/TROY-HA cells transfected for siJAK1. Cells were transfected with siRNA targeting JAK1 (siJAK1) or control siRNA (siCtrl) for 24 h. Cells were harvested and plated in DMEM with 0.1% serum into the top chamber of collagen-coated transwell inserts with DMEM containing 10% serum in the bottom chamber. Twenty-four hours later, the migrated cells were fixed and counted. Immunoblots show the knockdown efficiency of JAK1 siRNA. **, p < 0.01; ***, p < 0.001. (B) Immunoblots for the lysates of serum starved T98G/TROY-HA treated with increasing concentration of ruxolitinib (Ruxo) show potent inhibition of STAT3 activation. (C) Migration assay for T98G and T98G/TROY-HA cells treated with ruxolitinib. Serum starved T98G/TROY-HA cells were treated with ruxolitinib for 24 h. Cells were harvested and plated in DMEM with 0.1% serum into the top chamber of collagen-coated transwell inserts with DMEM containing 10% serum in the bottom chamber. Twenty-four hours later, the migrated cells were fixed and counted. T98G cells were used a control. Results indicate that increased TROY expression significantly increased cell migration which was inhibited by treatment with ruxolitinib (Ruxo). **, p < 0.01; ***, p < 0.001.