Additional file 5 of CX3CR1 deficiency aggravates amyloid driven neuronal pathology and cognitive decline in Alzheimer’s disease

Additional file 5: Supplemental Fig. 5. Flow cytometry based analysis of ex-vivo, microglial phagocytosis of fibrillar Aβ (fAβ). (A) Gating strategy to identify CD11b+ CD45low vs. CD11b+ CD45high microglia in methoxy-X04 injected 6 month-old cohorts. Flow cytometry plots shown represent data from B6;Cx3cr1+/+ and 5xFAD;Cx3cr1+/+ mice. (B) Flow cytometry plots showing the absence of non-specific retention of methoxy-X04 in CD11b+ microglia in B6;Cx3cr1+/+ mice and GFP+ microglia in B6;Cx3cr1-/- mice. (C) Flow cytometry data showing proportions of methoxy-X04+ microglia within CD11b+ CD45low and CD11b+ CD45high subpopulations in 6 month-old 5xFAD;Cx3cr1+/+ mice. Microglia identified using gating strategy showed in (A). fAβ+ microglia are further classified into methoxy-X04low / fAβlow and methoxy-X04high / fAβhigh sub-populations. (D) Flow cytometry data showing identification of microglia in 5xFAD;Cx3cr1-/- mice based on CD11b+ vs. GFP+ expression for analysis of proportion of phagocytic, methoxy-X04+ sub-populations. Data in B and D show that the use of CD11b vs. GFP does not alter the profiles of fAβ+ microglia, and there is no non-specific, spectral overlap between CD11b, GFP and methoxy-X04 channels. All data representative of flow-cytometry analyses done using n = 5 female and 5 male mice of each genotype and processed in a single experiment. All experiments done using appropriate single-colored compensation controls to eliminate spectral overlap.