Figure 3.

Pore-forming and cytotoxic activity of Hbl depending on heat treatment of the single components. The rHbl components were each set to 1.5 pmol/µL in elution buffer. These were heated individually or as a 1:1:1 mixture for 10 min in 20 µL samples and subsequently applied in the cell culture tests. The end concentrations in the PI influx test equaled 37.5 pmol/mL each. (A) PI influx after 10 min treatment of rHbl 1:1:1 with rising temperatures. (B) PI influx after incubation of each single rHbl component for 10 min at rising temperatures. Total fluorescence counts differ slightly from those in (A). Color scheme as depicted in (A). (C) Cytotoxic activity determined in WST–1 (water soluble tetrazolium) bioassays after 10 min treatment of each single rHbl component with rising temperatures.