Figure 6
From: Partial loss of Smad signaling during in vitroprogression of HPV16-immortalized human keratinocytes
TGF-β activation of a Smad-responsive luciferase reporter construct in HKc/HPV16 and HKc/DR. Four HKc/HPV16 and their corresponding HKc/DR lines (d-1, d-2, d-4 and d-5) were transiently transfected in triplicate wells per experimental condition with the p6SBE-Luc reporter construct and pRL-SV40. Cells were allowed to grow in CM for 24 h after transfections. Following this, they were treated without or with 40 pM TGF-B1 for 22 h and harvested for luciferase activity. Firefly luciferase values, measured in Relative Light Units (RLU), were normalized against Renilla luciferase values, and the results expressed as fold induction over control.