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. Author manuscript; available in PMC: 2016 Mar 1.
Published in final edited form as: J Biomol Screen. 2014 Nov 7;20(3):359–371. doi: 10.1177/1087057114557620

Figure 5. Confirming the effects of SID 7977862 from powder.

Figure 5

A) Structure of SID 7977862. B) Histograms of anti-LAMP fluorescence for unstimulated control (blue), stimulated control (red) and SID 7977862-treated cells. Experiment was conducted in NR. C) Dose-response testing using LAMP-1 externalization. Data are the average of 2 separate experiments conducted in NR+ 2% BSA. D) Assessing toxicity using Prestoblue reagent. Cells were pretreated with 100 μM SID 7877962 (triangle, 250,000 cells) or DMSO (circles, different numbers of cells), and Prestoblue fluorescence was measured. Different numbers of DMSO-treated cells treated with Triton X-100 (open circles) were used as an additional positive control. E) Testing for inhibition of calcium influx. F) Measuring effects on ERK phosphorylation. Experiments were conducted as in Figure 4A. G) Measuring effects on ERK catalytic activity. Experiments were conducted as in Figure 4C. H) Testing for inhibition of calcineurin. Experiments were conducted essentially as in Figure 4B.