Table 4.

Library of Pharmacologically Active Compounds and National Institutes of Health Clinical Collection pSTAT3 Primary High-Content Screening Summary

Compound library	No. of compounds	% of total
Compound information
LOPAC	1,280	74.2
NIH CC	446	25.8
Total	1,726	100
Cytotoxic and fluorescent outlier analysis
Ch 1: Average cell count/image <250	189	11.0
Ch 2: Hoechst average nuclear intensity z-score >3	8	0.5
Ch 2: Integrated inner intensity z-score >3	7	0.4
Ch 2: Average outer intensity z-score >3	15	0.9
Ch 2: Integrated outer intensity score >3	6	0.3
Primary HCS actives at 20 μM
% Inhibition of IL-6-induced pSTAT3 ≥50, 24 h exp.	51	3.0
Active confirmation at 20 μM
Mean % inhibition of IL-6-induced pSTAT3 ≥50, 24 h exp.	24	47.1
Mean % inhibition of IL-6-induced pSTAT3 ≥50, 1 h exp.	35	68.6
Mean % inhibition of IFNγ-induced pSTAT1 ≥50, 24 h exp.	6	11.8
Mean % inhibition of IFNγ induced pSTAT1 ≥50, 1 h exp.	18	35.3
Selective inhibitors of IL-6-induced pSTAT3 activation at 20 μM
Mean % pSTAT3 inhibition ≥50% (24 and 1 h) and mean % pSTAT1 inhibition <30% (24 and 1 h)	3	5.9

HCS data represent % inhibition of compounds tested in a single well at 20 μM, and confirmation data represent mean % inhibition±SD (n=3) of compounds tested in triplicate wells at 20 μM, after 1 or 24 h of compound exposures.

IFNγ, interferon-gamma; LOPAC, Library of Pharmacologically Active Compounds; NIH CC, National Institutes of Health Clinical Collection.