In vitro analysis of phagocytosis, antimicrobial activity, and gene-expression profiles of mRNA-electroporated granulocytes. (A) Phagocytosis capacity after incubation of p47^phox-deficient AR CGD granulocyte-rich apheresis cells either non-EP or p47^phox mRNA EP transfection corrected with inactivated AF488 fluorescent-labeled particles of S aureus (multiplicity of infection [MOI], 5) (n = 1-4). (B) Hyphal damage to A fumigatus expressed as the percentage of antifungal activity by HV granulocyte-rich apheresis product non-EP control or eGFP mRNA transfection control compared with X-CGD patient granulocyte-rich apheresis product non-EP (naive), eGFP mRNA EP transfection control, or gp91^phox mRNA transfection corrected (n = 6 replicates for each condition). ANOVA test with multiple comparisons; *P < .05. (C) Gene-expression profiles in mRNA-transfected granulocytes at 24 hours post-EP. Log₂ fold-change of expression for genes related to the response to exogenous mRNA for EP mRNA samples relative to naive samples for each 3 CGD patients (P1-3). Mean of 2 experimental replicates shown for P1 and P3 and a single analysis for P2. Genes with absolute log₂ fold-change value ≥2 are formally defined as true positives. ns, nonsignificant.