Fig. 8: Compound 9 inhibits the hemolytic activity of C+ E. faecalis by decreasing the amount of extractable CylLS” and reducing the level of cytolysin expression.
A Treatment of an inoculum (OD600 = 0.35) of washed E. faecalis cells with 100 μM inhibitor 9 ( + 9), in the presence of washed rabbit erythrocytes. The hemolytic activity decreased relative to untreated controls (− 9). In total, four biological replicates (n = 4) for each condition were performed. Error bars represent the standard deviation and are centered at the mean. B Assessment of the ability of inhibitor 9 to block the hemolytic activity of live E. faecalis over a range of inhibitor concentrations under conditions described for (A). EC50 was determined by fitting the data from two biological replicates (n = 2) with the dose-response function using Origin Pro 2024. Errors in the EC50 are the errors calculated during the non-linear concatenate fit. C LCMS analysis of the concentration of CylLS” extracted from overnight cultures of C+ E. faecalis ATCC 29212 treated with 9 or vehicle. CylLS” extract concentrations were determined by comparison to an internal standard of 15N,13C-labeled CylLS”. Four biological replicates were performed (n = 4). Errors bars are centered at the mean and represent the standard of deviation. D RT-qPCR data showing the amounts of cylLS, cylLL, and cylA transcripts extracted from overnight cultures of C+ E. faecalis treated with 100 μM of compound 9 relative to an untreated control (relative gene expression of 1.0). Four biological replicates were performed (n = 4). Error bars are centered at the mean and represent the standard of deviation. Statistical significance was calculated using a two sample and two sided t test for (A) and (C) and a one sample one sided t test for (D). **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file.
