Figure 27: . NF-κB translocation TNF-α stimuli Z-factor calculation & graph display.

Figure 27:

NF-κB translocation TNF-α stimuli Z-factor calculation & graph display. HeLa cells seeded at 5,000 cells/well overnight into 3 different 96-well plates were treated with 0.5% DMSO for maximum response (Red), media for unstimulated (green), and reference compound inhibitors IK202, 50µM for minimum response (blue) for 15 minutes followed by stimulation with 25ηg/ml TNF-α for ~35 minutes. Cells were then fixed and labeled to measure NF-κB translocation. Plates were analyzed on HCS imager to determine NF-κB translocation using CytoNuc Difference (Panel A) or Nuc/Cyt Ratio (Panel B) calculation. Raw data values from HCS instrument were used to calculate the mean response (left axis) and standard deviation (right axis). Z-factor was calculated as previously described.

From: Nuclear Factor Kappa B (NF-κB) Translocation Assay Development and Validation for High Content Screening

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Assay Guidance Manual [Internet].
Sittampalam GS, Coussens NP, Brimacombe K, et al., editors.
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