Fig. 5. Macrophages promoting HSC/MPP expansion.

a Immunofluorescence analysis showing the expression of F4/80 (representing macrophages) and c-Kit (representing HSCs/MPPs) in E14.5 FL cryosections. b Immunofluorescence analysis showing the expression of F4/80 (representing macrophages), and Runx1 (representing HSCs/MPPs) in E14.5 FL cryosections. c The proportion of HSCs/MPPs with and without cellular interaction, respectively (n > 3 biological replicates). d Immunofluorescence analysis showing the expression of F4/80 (representing macrophages), CD150 (representing HSCs/MPPs), Mecom (representing HSCs/MPPs) and Hlf (representing HSCs/MPPs) in E14.5 FL cryosections. e Schematic diagram of the procedures for HSC/MPP co-culture with macrophages. f The number of HSCs/MPPs (LSK cells) derived from macrophage and HSC/MPP co-culture system. Macrophages were a combination of F4/80^low and F4/80^high macrophages at 1:1 ratio. n = 3 biological replicates. g The number of HSCs/MPPs (LSK cells) derived from the MDK (50 ng/mL)-supplemented culture of 100 HSCs/MPPs (LSK, Flt3^–LSK and SLAM-LSK cells). n = 3 biological replicates. h Total cell number in E14.5 FL after Clodronate-liposome and Control-liposome treatment. Clodronate-liposome, Clodronate-L; Control-liposome, Control-L. n > 3 biological replicates. i The proportion of Lineage^– (Lin^–) cells in E14.5 FL after Clodronate-liposome and Control-liposome treatment. n > 3 biological replicates. j The proportion of LSK cells in E14.5 FL after Clodronate-liposome and Control-liposome treatment. n > 3 biological replicates. k The proportion of SLAM-LSK cells in E14.5 FL after Clodronate-liposome and Control-liposome treatment. n > 3 biological replicates. l Flow cytometry analysis showing the proportions of Lin^–, LSK, SLAM-LSK cells in E14.5 FL after Clodronate-liposome and Control-liposome treatment. Scale bars, 20 μm. The statistical results are presented as means ± SEM. Student’s t-test; *P < 0.05; **P < 0.01.