Figure 2. Doxycycline treatment drives a time-dependent increase in EmGFP expression in HeLa KRAS^G12V and BRAF^V600E clones expressing an EGR1:EmGFP reporter.

(A) Parental HeLa TetR KRAS^G12V cells (left) or HeLa TetR BRAF^V600E cells (right) along with clones expressing an EGR1:EmGFP reporter (KR1, KR2, KR4, KR8; BR1, BR2, BR4, BR5) were treated with 100 ng/ml Doxycycline (Dox) for between 0 and 72 h. Cells were fixed, stained with DAPI and imaged by high content microscopy. Results were analysed to determine the percentage of GFP-positive cells and mean GFP intensity, with GFP intensity from parental cells taken as background. Results represent the mean ± S.D. from three independent experiments, each performed in triplicate. (B) The cells described above (A) were treated as above and whole cell lysates were fractionated by SDS–PAGE and probed with the indicated antibodies. Results are representative of three independent experiments.