Fig. 3. The hypoxia PRE in CARTIV promotors is responsive and most potent when upstream of the minimal promotor.

a Schematic drawing showing the different hypoxia element locations relative to the mini TK minimal promotor tested. b Representative plots of HEK293T cells infected using lenti viral vectors with RFP670 under the control of the indicated CARTIV promotor and ZsGreen controlled by the ef1α core promotor. At 72 h following infection, the cells were placed under hypoxic conditions, incubated for 18 h, harvested and analyzed by flow cytometry. Data shown are ZsGreen-positive, single-discriminated, and DAPI-negative results. c Geometric mean of RFP670 in ZsGreen-positive cells normalized to unstimulated cells, showing the average of duplicates, error bars indicate standard deviation. A t test was performed to test for significance. Results are from one representative experiment of four performed.