a) hCMEC cytotoxicity calculated based on LDH release assay. Supernatant was collected from hCMECs infected with CJB111+pDC, CJB111ΔessC+pDC, and CJB111ΔessC+pDCessC at MOI 10, 4–5 hours post-infection. Percent cytotoxicity was calculated based on 0% (uninfected) and 100% (+lysis buffer) lysis controls. b) Contact-independent cytotoxicity in hCMECs during GBS infection using 0.4 μm polycarbonate transwells in which GBS and cells were separated by the porous membrane. Cytotoxicity was measured 24 hours later. Supernatant from hCMEC compartments were plated after the experiment to ensure no bacterial contamination. Statistics reflect the repeated-measures one way ANOVA with Tukey’s multiple comparisons, p < 0.05, *; p < 0.01, **. Data represent the mean of three independent experiments and error bars represent standard error of the mean.