mRNA expression analysis

Strain specific analysis of TPBG expression in WT and B-hTPBG mice by western blot. Spine were collected from WT mice and homozygous B-hTPBG (H/H) mice. Mouse TPBG was detectable in WT mice, the observed band size 63 kDa is the target band. Human TPBG was exclusively detectable in homozygous B-hTPBG and hTPBG overexpressed MC38 cells but not WT mice.

Protein expression analysis

Representative human TPBG expression in different tissues of B-hTPBG mice by IHC. Tissues were stained with antibodies specific for human TPBG (A-K) or anti-IgG antibody (L). Brain and spine of B-hTPBG mice show human TPBG positive. Human TPBG + cells were located in the white matter region of the spinal cord and the cortex region of the brain tissue, respectively. Human lung carcinoma as a positive control (J); Dog spleen was a negative control (K); Mouse lung tissues from the wild-type C57BL/6 mice (L).Other tissues were got from homozygous B-hTPBG mice (A-I). Original magnification ×200. Abbreviations: IHC, immunohistochemistry.

Analysis of leukocytes subpopulation in B-hTPBG mice

Blood routine test in B-hTPBG mice

Complete blood count (CBC). Blood from female C57BL/6 and B-hTPBG mice (n=6, 6-8 week-old) was collected and analyzed for CBC. Results indicate that introduction of hTPBG in place of its mouse counterpart does not change blood cell composition and morphology. Values are expressed as mean ± SEM.

Blood chemistry tests of B-hTPBG mice. Serum from the C57BL/6 and B-hTPBG mice (n=6, 6-8 week-old) was collected and analyzed. There was no differences on either measurement between C57BL/6 and B-hTPBG mice, indicating that introduction of hTPBG in place of its mouse counterpart does not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.