scMAYV-E immune sera protect monocyte derived macrophages (MDMs) from MAYV infection-induced cell death.

(A) Phase contrast images of MAYV-infected MDMs over time. Magnification: x20. (B) Phase contrast and fluorescent images of MDMs infected with MAYV preincubated with immune sera. 3 day-old cultures of MDMs were treated with DMEM media only, MAYV plus pVax1 sera, or MAYV preincubated with 1:100 dilution of pooled day 35 (post third immunization) immune sera from scMAYV-E immunized mice. After 48 hours of co-culture, the cells were fixed, permeabilized, and stained with Live Cell Labeling Kit-Green Fluorescence-Cytopainter, which stains only live cells with Labelling Dye Green. Magnification: ×40. (C) Histogram comparing the percentage of live cells in the control and experimental groups evaluated by Labelling Dye Green signals from six independent evaluations of the infected MDMs in Fig 3B. (D) Percent viability of Vero CCL-81 cells inoculated 36 hours with supernatants from the infected MDM cultures from experiments described in Fig 3B. The cell viabilities of Vero cells were assessed by Trypan Blue dye exclusion staining using a Countess II Automated Cell Counter. Each dot represents the cell viability from a single well +/- SEM counted in triplicates.