Fig. 3

Dual administration of low-dose allogeneic MSCs induces a higher proportion of anti-inflammatory mononuclear phagocytes in the draining lymph nodes. Draining lymph nodes (dLNs) were harvested 2 days post-transplantation (D+2) from corneal allograft recipient mice receiving either two injections of PBS or low-dose allogeneic MSCs the day before transplantation (D−1) and the day after transplantation (D+1). a Flow cytometry gating strategy used to select activated dendritic cells (DCs) (CD11c+MHCII+, CD11c+CD80+) or mononuclear phagocytes (MPh) with either a pro-inflammatory (CD11b+MHCII+, CD11b+CD80+) or an anti-inflammatory (CD11b+CD206+) phenotype. b Proportion of MHCII+ DCs expressed as a percentage of the parent (CD11c+) population. c Proportion of CD80+ DCs expressed as a percentage of the parent (CD11c+) population. d Proportion of MHCII+ MPh expressed as a percentage of the parent (CD11b+) population. e Proportion of CD80+ MPh expressed as a percentage of the parent (CD11b+) population. f Proportion of CD206+ MPh expressed as a percentage of the parent (CD11b+) population. g Analysis of mRNA expression (normalized to the housekeeping gene GAPDH and shown as fold-change relative to the PBS-treated allogeneic control group) in the dLNs of TGF-β1 at D+2 from PBS-treated allogeneic controls and low-dose allogeneic MSC-treated corneal allograft recipients. Error bars: mean ± SD. *p < 0.05 (each individual dot represents a separate animal, n = 3–6). D’Agostino and Pearson omnibus normality test and Shapiro-Wilk normality test used to determine the distribution of data. ROUT testing was used to identify outliers. Non-parametric unpaired two-tailed Student’s t tests used for data that was not normally distributed