Fig 2: Deletion of platelet Thbs1 in mice shields kidneys from fibrotic changes post‐I/R injury. Wild‐type or Thbs1 knockout mice were subjected to the surgery for bilateral I/R kidney injury or unilateral ureteral obstruction (UUO) and housed for 7 days. After that, mice were sacrificed, and kidney tissues were harvested for immunofluorescence. A) Representative images and the quantifications of THBS1/CD206/Ki67 immunofluorescence (IF) signal in the kidney (n = 5 for each group). Wild‐type or Thbs1 knockout mice were subjected to the surgery for bilateral I/R kidney injury or UUO and housed for 21 days and 14 days (n = 5 for each group), respectively. After that, all the mice first received Nuclear Magnetic Resonance Imaging (NMRI), followed by sacrificing, and kidney tissues were harvested for histopathology, Scale bar: 50 µm. B) Representative MASSON staining images and the quantifications of interstitial collagen accumulation (blue signal), Scale bar: 50 µm. C) Representative images and the quantifications of α‐SMA IHC signal in the kidney (n = 5 for each group). The whole pictures of the serial sections of the stained kidney were also displayed, and statistics between the indicated groups were analyzed using an unpaired two‐tailed Student's t‐test, and the significance was labeled with an asterisk; Scale bar: 50 µm. D) Representative NMRI images of the mouse kidney and the quantifications of T2 mapping for the kinetic motion of water molecules in the renal cortex; All values are presented as mean ± s.e.m.; n = 5 for each group, the statistics were analyzed using an unpaired two‐tailed Student's t‐test, * p < 0.05.

Fig 3: Platelet THBS1 antagonism diminishes cycling M2 proliferation and alleviates kidney fibrosis. A) Spatial feature plot of Thbs1 in ST spots of the IgG‐treated injured kidney as compared to the R300‐treated ones and Violin plot showing its relative expression. B) Feature plot showing the colocalization of Thbs1 and cycling M2 in ST spots of the IgG‐treated injured kidney and the R300‐treated ones. Mice received the kidney I/R injury treatment with or without THBS1 inhibitor LSKL. 7 or 21 days after the I/R injury, mice were sacrificed and kidney tissues were harvested for immunofluorescence (Day 7) or histopathology (Day 21), respectively. C,D) RAW 264.7 cells were treated with different doses of THBS1 and subjected to flow cytometry for Ki67 expression (C), or cell counting (D). E) Representative fluorescence images of CD206, Ki67, and THBS1 in the kidneys of the indicated three groups (n = 5 mice per group), Scale bar: 50 µm. F–H) The relative mRNA expression of Tnf, Il6, and Acta2 were analyzed in the kidneys of the indicated three groups (n = 5 mice per group). I) Representative whole kidney staining images of MASSON, and IHC for α‐SMA from each group were displayed (n = 5 for each group), Scale bar = 50 µm. All values are presented as mean ± s.e.m. The statistics were analyzed using an unpaired two‐tailed Student's t‐test, * p < 0.05.

Fig 4: The effect of PHLDA1 positive smooth muscle cells on head and neck squamous cell carcinoma in vitro.(A) Immunofluorescence analysis illustrating the expression profiles and localization of PHLDA1 in head and neck squamous cell carcinoma (HNSCC) and oral mucosa.(B, C) PCR analysis revealing differences in PHLDA1 gene and THBS1 gene transcription levels in HCASMC between the control and overexpression groups (***P < 0.001).(D, E) CCK-8 assay showing the effect of THBS1 on Cal27 cells and HN6 cells.(F) PCR analysis revealing differences in SDC1 gene transcription levels in Cal27 cells between the control and interference groups.(G) Western blot analysis showing differences in SDC1 protein levels in Cal27 cells between the control and interference groups.(H) Statistical significance of the migration area difference. (**P < 0.01, ***P < 0.001, ****P < 0.0001)(I) Statistical significance of the difference in migrating cells. (****P < 0.0001)(J) Wound healing assay demonstrating differences in migration area observed via microscopy.(K) Transwell assay illustrating the differences in cell numbers crossing the membrane.

Fig 5: Visual analysis of THBS signaling pathway.(A, B) The number (left) and strength (right) of cellular interactions circled plots with C4 PHLDA1+ SMCs as source (A) and tumor as target (B).(C) Heatmap demonstrated the centrality score of the THBS signaling pathway network, showing the relative importance of each cell group.(D) Heatmap showed the cell interactions of the THBS signaling pathway.(E, F) Violin and bubble plots demonstrated cellular interactions in the THBS signaling pathways.(G, H) Circle plot and hierarchical plot showed the inferred intercellular communication network for THBS1-SDC1 signaling. Solid and hollow circles indicated source and target cell types in hierarchical plot, respectively. The edge color of the middle circle was consistent with the signal source in hierarchical plot.