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. Author manuscript; available in PMC: 2014 May 1.
Published in final edited form as: J Biomol Screen. 2013 Nov 26;19(4):556–565. doi: 10.1177/1087057113513640

Figure 4.

Figure 4

Normalization of the Nef-BiFC signal to mRFP expression improves assay performance. Transfected 293T cells expressing the wild-type Nef-BiFC protein pair (WT2A) or the dimerization-defective control (4D2A) were imaged on the ArrayScan II platform, and the resulting data were analyzed in three different ways: (A) number of Nef-BiFC-positive (BiFC+) cells within each mRFP-positive cell population; (B) overall Nef-BiFC fluorescence intensity within the mRFP-positive subpopulation; (C) Nef-BiFC fluorescence intensity normalized to mRFP fluorescence intensity within each mRFP-positive cell. The statistical parameters resulting from each analysis are summarized in the tables on the right. Note that analysis based on the BiFC/mRFP ratio minimized variability and maintained high assay performance (Z factor > 0.6).