THBS1 Antibody - #DF6848

Fig. 5: Cell-to-cell communications among thyrocyte and immune cells predicted by the CellChat software in PTC and ATC. A Circle plot summarizing the maximum number of interactions among individual cell types in PTC. The thickness of the lines connecting cells indicates the interaction strength. B The same plot as in (A) but applied to ATC. C Differences of number and strength overall information flow within the inferred networks between PTC and ATC. D The potential outgoing and incoming interaction strength of each cellular cluster in PTC and ATC. Heatmap showing the relative signaling contribution of each cell group based on the number and strength compared PTC with ATC. E Heatmap showing the relative signaling contribution of each cell group based on the number and strength compared PTC with ATC. F Differential strength of network centrality based on APOE+ macrophages (PTC vs. ATC). G Bubble heatmap showing the cell–cell communication of selected ligand–receptor pairs between APOE+ macrophage and CD8+ PDCD1+ T cells. Dot size indicates P value, colored by communication probability. H Multiplex IHC shows the cross-talk between APOE+ macrophages and T cells via the ligand–receptor of THBS1-CD47.

Fig. 3. Fibrosis phenotype and THBS1/TGFβ1 expression were significantly elevated in the LFH group. (A) The thickness of LF was measured by MRI before operation. The thickness of LF at the L4/5 facet joint level was indicated by red arrow. (B) Representative tissues of LF acquired from patients with spinal surgery. The top three were from non-LFH patients and the bottom three were from LFH patients. Scale bar = 5 mm. (C) Representative images of H&E, MASSON, and EVG staining of the non-LFH group (n = 6) and the LFH group (n = 6). The yellow dotted area represents collagen fibers. Scale bar = 20 μm. (D) Immunohistochemical staining of THBS1, TGFβ1, COL1A2, and α-SMA in the LF samples from LFH group (n = 6) and non-LFH group (n = 6), and the percentage of positive cells was quantified. Scale bar = 20 μm. (E) TEM of LF ultrastructure of non-LFH and LFH groups. The yellow area represents elastin fibers. Scale bar = 8 μm. (F) The qRT-PCR analysis results showing that LFH group (n = 6) had a higher mRNA level of THBS1, TGFβ1, ACTA2 (α-SMA), and COL1A2 compared with non-LFH group (n = 6). The 2−ΔΔCt method was used to normalized with GAPDH. (G) Western blotting assay of THBS1, TGFβ1, COL1A2, and α-SMA protein levels of LF samples of the two groups. GAPDH served as the loading control. EVG: verhoeff's van gieson, TEM: transmission electron microscope, EF: elastic fiber, CF: collagen fiber. **p < 0.01, ***p < 0.001.

Fig. 7. The mechanical stress-THBS1 axis promotes hypertrophy and fibrosis of LF in vivo. (A) H&E staining of tissues from mice (n = 6 mice per group), and the quantitative analysis of LF area. Scale bar = 50 μm. (B) EVG staining of tissues from mice, and the quantitative analysis of volume fraction of elastin fibers in LF. Scale bar = 20 μm. (C, D) Immunohistochemical staining of α-SMA and THBS1 in mice tissues, and the percentage of positive cells was quantified. Scale bar = 20 μm. Error bars: mean ± S.D. ns, not significant. *p < 0.05, **p < 0.01, ***p < 0.001.

Figure 2. - Proteomic analysis following THBS1 overexpression. (A) Fluorescent imaging (×40; scale bar, 200 µm) of NCoe and THBS1oe groups after lentiviral-mediated THBS1 overexpression. (B) Evaluation of THBS1 overexpression using reverse transcription-quantitative PCR and western blot analysis. (C) Hierarchical clustering heatmap of differentially expressed proteins. (D) Volcano plot of DEPs: Green dots indicate downregulated proteins, and red dots indicate upregulated proteins. (E) GO and (F) KEGG pathway enrichment analysis of DEPs. **P

Figure 2. THBS1 expression levels in SGC tissues and cells. (A) Immunohistochemical staining of THBS1 in SGC and para-carcinoma tissues. Expression of THBS1 was negative in SGC tissues, but positive in para-carcinoma tissues. Scale bar, 50 µm. (B) Positive expression rate of THBS1 in eyelid SGC and para-carcinoma formalin-fixed paraffin-embedded samples. (C) Relative mRNA expression levels of THBS1 in SGC cells were lower than those in normal SG cells. **P<0.01 and ***P<0.001. SGC, sebaceous gland cancer; THBS1, thrombospondin 1.

Figure 4. miR-3907 negatively regulates THBS1 expression. (A) Predicted binding sites for miR-3907 within the wild-type and mutant 3'UTR of THBS1. (B) Quantitative analysis of the relative luciferase activity of cells with reporter vectors containing the wild type or mutant THBS1 3'UTR, following miR-3907 co-transfection. (C) Reverse transcription-quantitative PCR and (D) western blotting with semi-quantification analysis revealed decreased THBS1 expression following miR-3907 mimics transfection. *P<0.05 and ***P<0.001. miR, microRNA; THBS1, thrombospondin 1; UTR, untranslated region; NC, negative control.

Figure 5. Inhibitory effects of THBS1 on SGC cell proliferation and migration. (A) THBS1-knockdown was induced by transfecting SGC cells with siRNA-THBS1. THBS1 expression level was determined by reverse transcription-quantitative PCR. Results of (B) CCK-8 and (C) wound-healing assays after transfection with siRNA-THBS1; siRNA-THBS1 increased the proliferation and migration of SGC cells. (D) THBS1 overexpression was induced by transfecting lentiviral vectors; THBS1 expression level was determined by reverse transcription-quantitative PCR. (E) Fluorescence assay results revealed that the THBS1 lentivirus vector was successfully transfected. (F) CCK-8 and (G) wound-healing assays results after transfection with THBS1 lentivirus revealed that THBS1 decreased the proliferation and migration of SGC cells. Scale bar, 500 µm. *P<0.05, **P<0.01 and ***P<0.001. SGC, sebaceous gland cancer; siRNA, small interfering RNA; CCK-8, Cell Counting Kit-8; THBS1, thrombospondin 1; NC, negative control.