Fig. 8

Quantification of endogenous and injected macrophages in injured nerves following perineural transfer of CFSE-labeled polarized macrophages. a Representative dot blot of F4/80 and CFSE staining after one injection of DMEM only, showing endogenous macrophages (F4/80⁺CFSE⁻) in the top left quadrant, and the lack of injected macrophages (F4/80⁺CFSE⁺) in the top right quadrant. b Representative dot blot of F4/80 and CFSE staining after one injection of CFSE-labeled M2 cells, showing endogenous macrophages (F4/80⁺CFSE⁻) in the top left quadrant and injected macrophages (F4/80⁺CFSE⁺) in the top right quadrant. c Total number of endogenous macrophages (F4/80⁺CFSE⁻) after one injection of DMEM alone or CFSE-labeled M0, M1, and M2 cells. d Total number of endogenous macrophages (F4/80⁺CFSE⁻) after two injections of DMEM alone or CFSE-labeled M0, M1, and M2 cells. e Total number of injected macrophages (F4/80⁺CFSE⁺) after one injection of DMEM alone or CFSE-labeled M0, M1, and M2 cells. f Total number of injected macrophages (F4/80⁺CFSE⁺) after two injections of DMEM alone or CFSE-labeled M0, M1, and M2 cells. In all experiments, DMEM- or CFSE-labeled macrophages were injected at the injury site on day 14 (one injection) or on days 14 and 15 (two injections) after CCI. Animals were killed 24 h after the first injection and 24 h after the second injection, and cells from the nerves were analyzed using flow cytometry and FlowJo software. *p < 0.05, **p < 0.01, ***p < 0.001 (one-way ANOVA, Bonferroni’s test); ^## p < 0.01, ^### p < 0.001 compared to the same treatment 24 h after the first injection (unpaired t test). Data show the mean ± SEM (n = 6 animals per group)