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. 2013 Jun 1;6(3):382–391. doi: 10.1593/tlo.13232

Figure 1.

Figure 1

Optimization of a bioluminescent HTS to detect changes in N-linked glycosylation. (A) Schematic demonstrating the principle for signal detection using the ERLucT reporter vector: Luciferase is translated into the ER, glycosylated, and inactivated. However, when ER function is disrupted, the luciferase retains bioluminescent activity. (B) DMSO concentration-activity relationship in the D54-ERLucT cell line; representative experiment showing average of eight wells per DMSO concentration. (C) Cell density optimization for 96-well plate format using 1 µM of the GlcNac-phosphotransferase inhibitor Tn; representative experiment showing average of eight wells per treatment. (D) Dose response of Tn under optimized conditions. Data represent average of eight wells per Tn dose. Tn activates luciferase activity with a half-maximal effective concentration of 40 nM.