Fig. 4: Subcellular localization and characterization of selected mRNA VZV gE vaccine candidates in Vero cells.

VZV gE protein presence in the Golgi was assessed 24 h after transfection in Vero cells using spinning disk confocal microscopy. a DS from the three lead candidates: gE WT_Co2, gE_ms5 Co1 and gE_ms6 Co2 were transfected into Vero cells. 24 h later cells were washed, permeabilized and fixed. gE protein was detected using a human mAb against VZV gE and A488 secondary antibody. Golgi apparatus was stained using a specific antibody against N-acetylgalactosaminyltransferase 7 (GALNT7) and using A555 as a secondary antibody for detection. Nuclei were stained using DAPI (blue). Small rectangles represent the areas that were selected for analysis of colocalization. b A total of ten images were acquired per condition and colocalization levels between VZV gE protein and Golgi were measured for each mRNA construct using Pearson’s correlation coefficient based on regions of interest (ROI). One-way Analysis of Variance (ANOVA) with Tukey’s multiple comparison test was performed to determine statistical significance between the sample groups. ****p < 0.0001; ns: not significant.