TCL1 is the actor of cellular and biochemical phenotypes observed in Jurkat cells. (A) Jurkat-TCL1 cells were transduced with shRNA(GFP)-C and shRNA(GFP)-TCL1 vectors and stimulated with PMA (left) or UCHT1 (right) at the times indicated in the figure. ERK phosphorylation and TCL1 expression were determined by Western blotting. Total ERK expression was used as a loading control. A vertical line has been inserted to indicate a repositioned gel lane. A representative experiment of 2 performed is shown. (B) Photomicroscopy of Jurkat-TCL1 transduced with shRNA(GFP)-C and shRNA(GFP)-TCL1 vectors and treated or untreated with PMA for 3 days. (C) Jurkat cells were cotransfected with TCL1, MTCP1, or control vector and GFP-expression vector. Sorted GFP-positive transient transfected Jurkat cells (Jurkat-C*, Jurkat-TCL1*, and Jurkat-MTCP1*) were stimulated with PMA for 20 minutes (left). ERK phosphorylation and ERK expression were determined by Western blotting. After quantification, the p-ERK/total ERK ratio was determined as indicated at the bottom. (D) Transient transfected cells Jurkat-C* and Jurkat-TCL1* were stimulated with UCHT1. PKCθ phosphorylation and TCL1 expression were determined by Western blotting. Total PKCθ expression was used as a loading control.