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. 2023 Aug 3;13:12618. doi: 10.1038/s41598-023-39613-5

Figure 1.

Figure 1

MAC-dependent inner membrane permeabilization correlates with reduced viability of K. pneumoniae. (a) Survival on plate and inner membrane permeabilization of clinical K. pneumoniae isolates after 90-min incubation in 10% normal human serum (NHS) at 37 °C in the presence of 1 µM SYTOX green nucleic acid stain. Survival data was normalized to CFU counts in conditions where C5 conversion was inhibited by addition of 20 µg/ml OMCI and 20 µg/ml Eculizumab. Inner membrane permeabilization (SYTOX fluorescence intensity) was determined in a microplate reader. Red dotted line indicates background permeabilization signal (OMCI + Eculizumab). (b) Inner membrane permeabilization of Kp570, Kp702 and Kp193 in the presence of 10% NHS, 10% NHS in which C5 conversion was inhibited by addition of 20 µg/ml OMCI and 20 µg/ml Eculizumab (C5 inhibition), or 10% heat inactivated NHS (HiNHS). Bacteria were incubated at 37 °C in the presence of 1 µM SYTOX green nucleic acid stain, and inner membrane permeabilization (SYTOX fluorescence intensity) was detected every 2 min for 90 min in a microplate reader. (a,b) Data represent mean ± standard deviation of three independent experiments.