Fig. 7

Sparing of adverse effects on mammalian cells by CMLD013075. a Cultures of a mouse macrophage cell line were pre-incubated with the indicated concentrations of Hsp90 inhibitors, washed into compound-free medium and infected with a strain of GFP-expressing C. albicans. After an additional 2-h incubation, engulfment of fungus was monitored by flow cytometry. The percentage of macrophages with internalized C. albicans is plotted relative to no-compound control for increasing concentrations of inhibitor. Each data point represents the mean of triplicate determinations. Error bars, SD. The experiment was repeated as an independent biological replicate to confirm results. b Effect of Hsp90 inhibitor exposure on fungal filamentation, a well-established consequence of Hsp90 inhibition was imaged by differential interference contrast microscopy. Representative fields from micrographs obtained at the same magnification for all images are presented. Scale bar, 10 µm. The experiment was repeated as an independent biological replicate to confirm results. c Cultures of a mouse macrophage cell line were pre-incubated with the indicated concentrations of Hsp90 inhibitors, washed into compound-free medium and infected with a strain of GFP-expressing C. albicans. After additional 2-h incubation, aliquots of medium were removed and assayed by ELISA for level of the indicated pro-inflammatory cytokines. Each data point depicts the mean derived from three independent cultures. Error bars, SD. The entire experiment was repeated as an independent biological replicate to confirm results. d Induction of the heat-shock response was measured in a strain of C. albicans (right) and a mammalian cell line (left). Cells of each species were stably transduced with a plasmid encoding a reporter protein under transcriptional control of heat-shock promoter elements from an HSP70 gene native to the respective species. Relative concentration-dependent activation of reporter expression by compounds after overnight (human cells) or 6-h (C. albicans) exposure is plotted. Each point depicts the mean of three independent wells. Error bars, SD. The experiment in both human and fungal cells was repeated as an independent biological replicate to confirm results