Fig. 3

The role of TIMELESS on the activation of NF-κB signal pathway. a ZR-75-30 cells were transfected with NF-κB luc and either with p65 or TIMELESS, p65 and TIMELESS for 24 h and then the luciferase activity was measured (up). The cell lysate was subjected to western blot with anti-GFP or anti-Flag antibody (down). b ZR-75-30 cells were transfected with control vector or TIMELESS and then subjected to the real-time PCR analysis after 24 h transfection. c The schematic of MMP9-Luc, NF-κB del MMP9-Luc, and NF-κB mut MMP9-Luc. d ZR-75-30 cells were transfected with MMP9-luc, NF-κB del MMP9-Luc or NF-κB mut MMP9-Luc together with overexpression of TIMELESS or not for 24 h. After that the luciferase activity was measured (up). The cell lysate was subjected to western blot with anti-Flag antibody (down). Each bar represents the mean ± s.d. from three independent experiments. “*”, p < 0.05; “**”, p < 0.01