The SUMO pathway regulated the protein levels of DAPK1. HEK293T were transfected with (A) HA-DAPK1, LacZ, V5-UBC9, His-SUMO-1, His-SUMO-2 or His-SUMO-3, or (B) HA-DAPK1, LacZ, V5-UBC9, His-SUMO-1, His-SUMO-2 or His-SUMO-3, or (C) HA-DAPK1, LacZ and six different SENPs, or (E) control and UBC9 siRNA as indicated. HCT116 were transfected with (D) HA-DAPK1, LacZ and six different SENPs as indicated. Cell lysates were extracted 48 h post-transfection and immunoblotted with respective antibodies. The intensity of the bands was quantified using LICOR Odyssey software and represented by bar graphs. The experiments were repeated three times (n = 3) and representative images are presented. The representative western blot images are from different gels and each lane was loaded with the same amount of proteins. Data of triplicate assays are presented as mean ± S.D. * p < 0.05; ** p < 0.01; *** p < 0.001; NS, no significance. “+” indicated that the plasmid was transfected, “−” indicated that the plasmid was not transfected.