Fig. 1. Infection of human RBCs with V. vulnificus induces hemolysis and shape change.

a, b RBCs were infected with V. vulnificus at various MOIs and incubated for 1 h (a) (n = 5, two-tailed Student’s t-test) or for different times (b) (n = 6, two-way analysis of variance (ANOVA) followed by Duncan’s multiple range test) as indicated. c RBCs were infected with V. vulnificus at various MOIs and incubated for 1 h and shape changes were analyzed by SEM. The red arrowhead indicates V. vulnificus; white arrow, echinocyte; and the white arrowhead, spherocyte. The scale bar is presented. d Different shapes of RBCs were quantified from the SEM images obtained from six independent experiments and were expressed as the percent of the specific shapes per total RBCs (n = 6, two-tailed Student’s t-test). e RBCs were infected with V. vulnificus at various MOIs and incubated for 1 h. The representative images by confocal microscopy among five independent experiments were presented. Different shapes of RBCs are marked with the same symbols used for Fig. 1c. Scale bar is presented. f A schematic diagram represents a typical morphological change of RBCs under external stress. The means ± SE were calculated from at least five independent experiments. Control, uninfected; MOI multiplicity of infection, SEM scanning electron microscopy, and MV microvesicle.