JAK3/STAT5 blockade inhibits constitutive STAT5 tyrosine phosphorylation and BCL10/NFκB activity in MT-2 cells. (A) MT-2 cells were treated with medium, DMSO (0.1%, vehicle control) ascending amounts of NC1153 as indicated for 24 h. Total cell lysates were Western blotted with antibodies as indicated to the right. Molecular weight markers are indicated to the left. (B) JAK3 blockade diminishes BCL10-SBR occupancy by STAT5 in vivo. MT-2 cells were treated with NC1153 as indicated for 10 h. ChIP assays were performed with anti-STAT5 antibody and IgG control from each treatment. STAT5 DNA binding is expressed as fold change over background (IgG control). (C) NC1153 reduces NFκB activity and BCL10 protein expression. MT-2 cells were treated as described above for 48 h. Total cell lysates were Western blotted with antibodies indicated to the right. Molecular weight markers are shown to the left.