Rag2/IL2rg/Flt3-KO(BALB/c)

Fig.1 Percentage of T, B, NK cell subsets in spleen (n=5). 

Fig.2 Percentage of T, B, NK cell subsets in PB (n=5). 

Fig.3 Detection of mouse Flt3 Ligand expression in serum by ELISA (n=5). 

Abbr.  HO, homozygous; WT, wild type; N.D. not detected.

Fig.4 Body weight growth curve of M-NSG and Rag2/IL2rg/Flt3-KO (BALB/c) mice post human HSC inoculation. 

Human CD34+ HSCs (1E5) were intravenously injected into homozygous Rag2/IL2rg/Flt3-KO (BALB/c) and M-NSG mice (n=8-10, female, 3-4 weeks). 

Fig.5 Humanized immune reconstitution levels in M-NSG and Rag2/IL2rg/Flt3-KO (BALB/c) mice engrafted with human HSC. 

Human CD34+ HSCs (1E5) were intravenously injected into homozygous Rag2/IL2rg/Flt3-KO (BALB/c) and M-NSG mice (n=8-10, female, 3-4 weeks). All mice received 1.0 Gy irradiation prior to cell inoculation. Peripheral blood were analyzed by flow cytometry at indicated time points post inoculation. No significant differences were observed between the two mouse strains across the examined human immune cell subsets.

Fig.6 Percentages of pDCs, cDCs, and monocytes within human CD45 population were determined in the spleen of Rag2/IL2rg/Flt3-KO (BALB/c) mice by FACS. 

After 16 weeks of human immune reconstitution, mice received intraperitoneal injections of hFlt3L‑Fc (5 μg per mouse) three times per week for 2 weeks. One week after the final injection, mice were euthanized and spleens were harvested for immune cell analysis. The results demonstrated that hFlt3L‑Fc treatment significantly increased the percentages of plasmacytoid dendritic cells (pDCs) and conventional dendritic cells (cDCs), while moderately elevating the proportion of monocytes (n=4–5 females per group) (Mean±SEM, Unpaired t-test, ***P< 0.001).