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. 2019 May 23;63(6):e00342-19. doi: 10.1128/AAC.00342-19

FIG 4.

FIG 4

Lysocin cytotoxicity toward eukaryotic cells and bacterial endotoxin release. (A) hRBCs were incubated in triplicate with buffer or 0.5 to 256 μg/ml PyS2-GN4 for 8 h at 37°C, and hemolysis, as a function of hemoglobin release, was assayed spectrophotometrically at 405 nm. Triton X-100 was used as a positive control for hemolysis. (B) Human promyeloblast HL-60 cells were incubated in triplicate with buffer or 0.5 to 256 μg/ml PyS2-GN4 for 8 h at 37°C, and viability, as a function of formazan product formation, was measured spectrophotometrically at 570 nm. Triton X-100 was used as a control for cytotoxicity. (C) Endotoxin release was measured in duplicate experiments after treating P. aeruginosa strain 453 at 106 CFU/ml for 1 or 4 h at 37°C in growth medium with 0.2× and 5× MIC of PyS2-GN4, colistin, meropenem, or tobramycin. An untreated control was included. All error bars correspond to ±SEM. P values were calculated using a one-way analysis of variance. EU, endotoxin units.