Fig 2: Differences in TIM-3 expression between LEAOD and normal arterial tissues. (A) Immunohistochemical analysis of TIM-3 expression in LEAOD and normal arterial tissues (magnification, ×400). (B) TIM-2 and PDGF-BB protein expression levels in LEAOD and normal arterial tissues were determined by western blotting and quantification. (C) Correlation between the relative expression of PDGF-BB and TIM-3. *P<0.05, **P<0.01, ***P<0.001. TIM-3, T-cell immunoglobulin and mucin domain 3; LEAOD, lower extremity arteriosclerosis obliterans disease; PDGF-BB, platelet-derived growth factor-BB.

Fig 3: TIM-3 inhibits PDGF-BB-induced proliferation and migration in HASMCs. (A) Quantification of flow cytometric analysis of the proportion of TIM-3+ PDGF-BB-stimulated HASMCs. **P<0.01 and ***P<0.001 vs. control group (B) EdU, (C) Transwell and (D) wound healing assays were used to assess proliferation and migration in PDGF-BB-stimulated (20 ng/ml) HASMCs in the presence or absence of TIM-3 (1,000 ng/ml) and anti-TIM-3 (10 µg/ml; n=5; scale bar, 100 µm). *P<0.05, **P<0.01. TIM-3, T-cell immunoglobulin and mucin domain 3; PDGF-BB, platelet-derived growth factor-BB; HASMCs, human artery vascular smooth muscle cells.

Fig 4: IL-6 and TNF-α expression in the four treatment groups and the effect of TIM-3 overexpression on the NF-κB signaling pathway in PDGF-BB-stimulated HASMCs. Protein expression levels of (A) IL-6, (B) TNF-α, (C) TIM-3, p-p65 NF-κB and p65 NF-κB were detected by western blotting. *P<0.05, **P<0.01. IL-6, interleukin-6; TNF-α, tumor necrosis factor-α; TIM-3, T-cell immunoglobulin and mucin domain 3; PDGF-BB, platelet-derived growth factor-BB; HASMCs, human artery vascular smooth muscle cells; p, phosphorylated; LV, lentivirus.