Screening for compounds that reduce the HMGB1-induced pro-inflammatory response.

<p><b>A:</b> Timeline of the cell-based screening: RAW 264.7 cells were pre-treated with test compounds and exposed to HMGB1 for 18 hours. TNFα production was measured from the supernatant and the viability of the cells was measured by the MTT assay. <b>B:</b> Dot graph showing the individual TNFα/viability results of the tested 5,646 compounds. TNFα responses are shown as % values of the HMGB1-induced TNFα production. Values lower than MEAN-2SD are shown in red (viability) and green (TNFα response) boxes to denote “toxic” and “Hit” compounds. <b>C–D:</b> Distribution of viability (C) and TNFα response (D) data with superimposed Gaussian distribution curves fitted to the data points.