Fig1. Detection of TLR8 expression in spleen, lung and thymus by RT-PCR.?

Wild type: only one band at 318 bp with primers F1/R1(mTlr8);

Homoygous: only one band at 366 bp with primers F2/R2(hTLR8);?

Fig 2. The representative picture of hepatomegaly(top), splenomegaly (bottom) in aged hTLR8 mice.

?

Fig 3. Histology of (A) spleen and (B) liver in aged hTLR8 mice.?A prominent portal inflammation with fibrosis and bile duct hyperplasia in liver (A); Decreased cellularity in periarteriolar lymphoid sheath and the increase of histiocytes in the red pulp of spleen (B).

Fig 4. The expression pattern of hTLR8 in myeloid cells (A, B); and cytokine analysis in serum of aged hTLR8 mice (C).

Fig 5. The dynamic change of (A) myeloid and (B) lymphocyte lineage in the spleen of hTLR8 mice; and (C) in vitro stimulation of splenocytes from wild type (WT) C57BL/6 and hTLR8 mice with hTLR8 agonist ORN-06.

Fig 6. MC-38 tumor growth curve and tumor weight of individual mice from the 4 groups.?

One way ANOVA *, **, and *** refer to P < 0.05, P < 0.01, and P < 0.001, respectively.

Fig 7. Serum level of cytokine analysis by MSD and intracellular TNF-a staining by FACS in splenocytes.

Fig 8. Tumor infiltrated immune component analysis.

Fig 9. Pro-inflammatory cytokines in the tumor cell lysate.?*Validation data are provided by Crownbio.

Fig 10. No abnormalities were found in the alive mice until 29.1 weeks old.

* G2 #264 (HO hTLR8, male) died at 28 weeks old, with no sweeling in its limbs.

Fig 11. No abnormalities were found in the limbs, pancreas, liver and kidney. The spleens of hTLR8 mice have abnormally enlarged.

表1 評分標準

表2 各組織H＆E染色切片評分結果匯總

Fig.12 Partial representative pictures of bone pathology (29 weeks old Magnification: 100x).