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. 2014 Feb 1;12(1):55–79. doi: 10.1089/adt.2013.524

Table 3.

Three-Day 5-Plate Dimethyl Sulfoxide Validation of the IL-6–Induced pSTAT3-Y705 Activation High-Content Screening Assay

  Plate controls average inner intensity pSTAT3-Y705 Ch 2 Plate performance statistics
Plate ID Mean_Max (n=32) Mean_Min (n=32) Z′-factor S:B ratio
Day 1
 1 8,526.2 1,111.5 0.53 7.67
 2 8,444.2 1,113.7 0.41 7.58
 3 6,887.6 1,104.6 0.21 6.24
 4 8,444.2 1,188.2 0.53 7.11
 5 7,282.1 1,169.3 0.29 6.23
Day 2
 1 7,797.2 1,519.8 0.62 5.13
 2 7,767.1 1,372.3 0.59 5.66
 3 7,843.9 1,378.5 0.69 5.69
 4 8,401.7 1,481.0 0.62 5.67
 5 9,203.9 1,558.2 0.62 5.91
Day 3
 1 8,466.3 1,388.0 0.64 6.10
 2 7,889.1 1,368.2 0.53 5.77
 3 7,893.8 1,381.0 0.57 5.72
 4 8,020.5 1,351.9 0.55 5.93
 5 9,018.4 1,553.3 0.67 5.81

 The average inner intensity pSTAT3-Y705 Ch 2 data presented in the table were generated in three independent experiments of 5×384-well DMSO validation plates. These experiments were designed to mimic 3 days of automated screening operations in the pSTAT3-Y705 activation HCS assay, save that DMSO was added to wells instead of compounds. The average inner intensity pSTAT3-Y705 Ch 2 data of the maximum (50 ng/mL IL-6 and 0.2% DMSO, n=32) and minimum (0.2% DMSO, n=32) plate controls from each of the 15×384-well plates are presented.