Decreased TCL1 expression in SUP-T11 cell line restores PKCθ pathway activation. (A) SUPT-11 cells were transduced with shRNA-C, shRNA-TCL1(1), and shRNA-TCL1(4) vectors and stimulated with PMA (10 ng/mL) at the times indicated in the figure. ERK phosphorylation and TCL1 expression were determined by Western blotting. Total ERK expression was used as loading control. A representative experiment of 3 performed is shown. After quantification, the p-ERK/ERK ratio (top panel, white box: t = 0 minute; gray box: t = 5 minutes; black box, t = 30 minutes of PMA induction) and TCL1 expression inhibition (bottom panel, a white box represents the mean plus or minus SE of the 3 time points for each shRNA) were determined and related to shRNA control and represented as histograms. A representative experiment of 2 performed is shown. (B) SUPT-11 cells were transduced with shRNA-C, shRNA-TCL1(1), and shRNA-TCL1(3) vectors and stimulated with UCHT1 at the times indicated in the figure. PKCθ phosphorylation and TCL1 expression were determined by Western blotting. β-Actin expression was used as loading control. After quantification, the pPKCθ/β-actin ratio (top panel, white box: t = 0 minute; gray box: t = 5 minutes; black box, t = 30 minutes of PMA induction) and TCL1 expression inhibition (bottom panel, a white box represents the mean plus or minus SE of the 3 time points for each shRNA) were determined and related to shRNA control and represented as histograms. A representative experiment of 2 performed is shown.