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. Author manuscript; available in PMC: 2015 May 18.
Published in final edited form as: J Biomol Screen. 2012 Dec 10;18(6):647–658. doi: 10.1177/1087057112469405

Figure 1.

Figure 1

(A) Mutation of GLuc to eGLuc2 results in a more stable luminescent signal. HEK-293T cells were transfected with the indicated constructs, and conditioned media aliquots were assayed for GLuc activity. (B) Expression of eGLuc2 genes is tightly regulated and strongly inducible by doxycycline (dox) in Tet-On ARPE19 cells. Cells were cultured for 4 days in the presence (+) or absence (−) of 1 μg/mL dox, followed by a 24-h media change. eGLuc2 activity in the media after a total of 5 days expression was measured by the GLuc assay (n = 3, biological triplicates, ± SD). (C, D) Representative raw data from screening one 384-well Library of Pharmacologically Active Compounds (LOPAC) plate against cells expressing (C) wild-type (WT) fibulin-3–eGLuc2 (WT-eGLuc2) or (D) R345W fibulin-3–eGLuc2 (R345W-eGLuc2). The average vehicle-treated control signal + 3 SD (green dashed line) or – 3 SD (red dashed line) is shown for each cell line. (E) The two fibulin-3–enhancing compounds and top 10 fibulin-3–reducing compounds are displayed (n = 3, ± SEM).