hTFR1

Fig.1 Detection of TFR1 expression in liver by RT-PCR. 

Wild type: only one band at 243 bp with primers F1/R1(mTfr1);

Homozygous:  only one band at 279 bp with primers F2/R2(hTFR1).

Abbr. M, DNA marker; HO, homozygous; WT, wild type.

Fig2. Detection of human TFR1 expression in hTFR1 mice by WB. 

Abbr. HO, homozygous; WT, wild type.

Fig3. Expression characterization of human TFR1 on activated T cells in spleen in hTFR1 knockin mice.

Fig4. Detection of human TFR1 expression on erythroid cells in hTFR1 knockin mice-derived bone marrow. 

Fig5. Pattern of expression of activation markers overtime on activated CD4⁺ and CD8⁺ T cells upon anti-mCD3/mCD28 treatment. 

Note: The splenocytes were stimulated with anti-mCD3/mCD28 antibody in vitro.

Fig6. Detection of hTFR1 expression on endothelial cells in hTFR1 knokcin mice by immunofluorescence staining. 

Fig7. In vivo evaluation of TFR1-targeting therapeutic agents in hTFR1 knockin mice. (In cooperation with a partner)

In vivo knock-down efficiency of hTFR1-targted siRNA in liver and heart in hTFR1 knockin mice. 

Fig8. In vivo evaluation of TFR1-targeting therapeutic agents in hTFR1 knockin mice. (In cooperation with a partner)

hTFR1/CD71 knockin mice were injected with a control antibody (10mg/kg) and an antibody (10 mg/kg) against human TFR1 via i.v. injection. Brain tissues and serums were taken after 18 hr post administration. Brain concentrations, serum concentrations and brain-to-serum ratio of antibodies were quantified. As shown in the figure, hTfr binding antibody exhibited higher serum clearance and enhanced brain exposure. 

Fig.9 Blood routine test of hTFR1 mice (n=5 males and 5 females). Compared to WT C57BL/6 mice, a few parameters in the complete blood count (CBC) showed abnormalities.

Fig.10 Blood Biochemistry of hTFR1 mice (n=5 males and 5 females).

Fig.11 In vivo efficacy of an hTFR1-targeting antibody in hTFR1 knockin mice. The results demonstrated that hTFR1 mice exhibited enhanced serum clearance coupled with significantly elevated brain exposure, confirming efficient BBB penetration. Mice were administered a single intravenous (i.v.) injection of positive control antibody or hTFR-targeted antibody at a dose of 10 mg/kg. Serum was collected at 4, 8, and 24 hours post-dose. At 24 hours after injection, whole brains and brain parenchyma were harvested, and antibody concentrations in all samples were quantified using ELISA (n=3 females, 8 weeks old).