Recipient EIMs persist and expand after transplantation. (A-E) Representative flow cytometry of EIMs in naive (no-Tx) and posttransplant (5 weeks) MacGreen BM. (A and B) F4/80⁺Ly6G⁺ EIMs (gate i) were further segregated into naive and posttransplant recipient VCAM-1⁺GFP⁺ cells (gate ii) or posttransplant donor VCAM-1⁺GFP^neg cells (gate iii). (C-E) Representative flow histograms showing CD169, ER-HR3, MerTK, and TIM-4 expression in no-transplant (C), donor (D), and recipient (E) EIMs. Isotype control staining is represented by the blue line, and specific staining represented as the red line. (F) Total number of recipient (green bars) and donor EIMs (white bars) per femur across the transplantation time course (n = 5-13 mice/time point). Data are presented as mean ± SD. Statistical analysis was performed using a 1-way ANOVA Tukey’s multiple comparison test on recipient EIMs in transplanted groups, where asterisks indicate significant differences when compared with 2 weeks posttransplant (*P < .05 and *** P < .001). Flow cytometry plots and histograms as well as associated gated cell percent frequencies are from a representative animal, and the percentage of positive cells is based on the preceding parent populations. Data are representative of 5 to 13 individual animal samples per time point from either 1 (weeks 3 and 32), 2 (week 5), or 3 (all other time points) biological replicate experiments. 300 000 events were collected from each experiment sample by flow cytometry.