Figure 3.

EBP2 stabilized c-Myc. (a) HA-c-Myc was expressed in HEK293T cells with vector or an increasing dose of FLAG-EBP2. The protein levels of c-Myc were analyzed using western blotting. (b) HA-c-Myc was co-expressed with or without EBP2 in HEK293T cells. Cells were then treated with CHX for the indicated time. The expression of c-Myc, EBP2, and actin were determined using western blotting. (c) A549 cells or H1299 cells were transfected with siNC or two siRNAs against EBP2: siEBP2-1 (siE1), siEBP2-2 (siE2) for 72 h. The protein levels of c-Myc, EBP2, β-catenin, IκB-α and actin were determined using western blotting. The relative levels of c-Myc and EBP2 were quantified using Image J. (d) A549 cells were transfected with siNC, siEBP2-1, and siEBP2-2 for 72 h. Cells were then treated with CHX for indicated times. The expression of c-Myc, EBP2, and actin were determined using western blotting. The relative level of c-Myc was quantified by Image J. (e) A549 cells were transfected with siNC, siEBP2-1 and siEBP2-2 for 72 h. Cells were divided into two wells and one well was treated with proteasome inhibitor MG132 (10 μM) or DMSO as control for 8 h before collection