Fig. 2.

Design of experiment to optimize experimental assay conditions for the screen. (A) HeLa cell density growth kinetics at three seeding densities of 250, 500, and 1,000 cells per well, (B) dose–response studies of HeLa cells tolerance to polybrene addition, (C) dose–response studies of puromycin kill curve of HeLa cells estimating the IC₅₀ and IC₉₅ values, and (D) control shRNA hairpin transduction experiments using HeLa cell density of 500 cells per well in 8 μg/mL polybrene. Experiments were conducted by seeding HeLa cells as indicated in 384-well microtiter tissue-treated plates and monitoring their growth over a course of up to 10 days as indicated. Cell fixing, imaging, and nuclei quantification were performed as described in Material and Methods.