FIG 13.

HCMV infection reduces stimulus-evoked Ca²⁺ responses in organoid-derived neural cells. (A) Day 30 organoids were mock infected or infected using HCMV TB40/E-eGFP at 0.5 IU/cell and treated with DMSO vehicle control (+vehicle) or 10 μM maribavir (+MBV). Representative bright-field and fluorescent images of dissociated neurons and astrocytes were taken at 14 days postplating. (B) The mean GFP fluorescence decreased in HCMV-infected cells with MBV cotreatment. (C) Intracellular Ca²⁺ levels were measured at 14 days postplating with HCMV-infected cells separated into GFP-positive (G+) and -negative (G−) populations. Intracellular Ca²⁺ levels were measured prior to stimulation (left panel), and the total number of cells responding to 10 μM ATP stimulation (middle panel) and the percent Ca²⁺ response over baseline are shown (right panel). (D) Stimulation using 50 μM KCl as described for panel C. (E) Day 60 organoids were mock treated or infected as described for panel A. Intracellular Ca²⁺ levels were measured at 14 days postplating prior to stimulation (left panel), and the total number of cells responding to 10 μM ATP stimulation (middle panel) and the percent Ca²⁺ response over baseline are shown (right panel). (F) Stimulation using 50 μM KCl as described for panel E. Data were collected from four biological replicate experiments with 100 cells analyzed in each replicate. (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant, as determined by ANOVA or chi-square test).