Figure 2.

Effects of BFstatin on the BrpR activity, bacterial growth, and human cell viability. (A) The chemical structure of BFstatin, N1-(2-chloro-5-fluorophenyl)-N3-propylmalonamide. (B) The EC₅₀ of BFstatin inhibiting the relative BrpR activity (%) was calculated as described in the Materials and methods section. (C) Growth of the V. vulnificus strains along with either 20 or 100 μM BFstatin or 2% DMSO (control) was monitored at 2 h intervals using a microplate reader and expressed as A₆₀₀. WT, V. vulnificus CMCP6; ΔbrpR, V. vulnificus CMCP6 brpR mutant. (D) The relative cytotoxicity (%) of BFstatin was determined using LDH activities released from HeLa cells incubated at 37°C for 3 h with either 20 or 100 μM BFstatin or 2% DMSO (control). The cytotoxicity was expressed using the LDH activity from the cells completely lysed by 4% Triton X-100 as 100%. ND, not detected. Error bars represent the SD from biological triplicates (B,C) or the representative of three independent experiments (D).