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Figure 4 | BMC Biotechnology

Figure 4

From: The cumate gene-switch: a system for regulated expression in mammalian cells

Figure 4

A. Dose-dependent control of reporter gene expression by cumate: 293CymR-CMV5-CuO-GFP and 293cTA-CR5-GFP cells that were cultured under conditions where reporter gene expression was off (293CymR-CMV5-CuO-GFP in the absence of cumate and 293cTA-CR5-GFP, in the presence of 50 μg/ml cumate) were washed with PBS and incubated for 48 h in the presence of varying concentrations of cumate. Total GFP fluorescence was measured by flow cytometry and is indicated in the y-axis in terms of fluorescence index (% GFP positive cells X mean fluorescence index). The figure represents the data from triplicate measurements. B. Tight control of GFP expression in cells stably expressing CymR and pCMV5-CuO-GFP: A clone of 293CymR-CMV5-CuO-GFP was cultured for 48 h in the presence of various concentrations of cumate. The figure shows representative fluorescent images of the cells treated with 0, 3 and 30 μg/ml of cumate from the experiment shown in the panel A. C. Tight control of GFP expression in cells stably expressing cTA and pCR5-GFP: The enriched pool of 293cTA-CR5-GFP, that was cultured in the presence of cumate, was washed in PBS and incubated for 48 h in the presence of various concentrations of cumate. The figure shows representative fluorescent images of the cells treated with 0, 0.5 and 10 μg/ml of cumate from the experiment shown in the panel A.

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