Fig. 2

The effect of TIMELESS on the expression of MMP9. a The cells were transfected with MMP9 luc with or without Flag-TIMELESS, and then cells were collected and luciferase activity was measured after 24 h transfection. b Similar experiment as A was performed but with an increasing dose of Flag-TIMELESS. c The breast cancer cells were transfected with a control vector or Flag-TIMELESS for 24 h and then subjected to the real-time PCR analysis. d T47D cells were transfected with shcontrol vector or shTIMELESS#1 for 24 h, and then the mRNA level was measured by real-time PCR. e ZR-75–30 cells were transfected with control vector or Flag-TIMELESS for 24 h, followed by western blot with anti-MMP9, anti-Flag or anti-β-Tubulin antibodies. f MCF-7 cells were transfected with shcontrol vector or shTIMELESS#1 for 24 h, followed by western blot with anti-MMP9, anti-TIMELESS or anti-β-Tubulin antibodies. Each bar represents the mean ± s.d. from three independent experiments. “*”, p < 0.05; “**”, p < 0.01