Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: The therapeutic effect of THBS4-rich-PMSC-Exos in preeclamptic rats. ( A ) Schematic of our animal experiments. ( B ) Detection of sperm in vaginal smears on gestational day 1 (GD1) under a light microscope. ( C ) Location of placental injection (indicated by arrow). ( D and E ) Systolic blood pressure (SBP) measurements and urinary protein levels on GD15. ( F and G ) Systolic blood pressure (SBP) measurements and urinary protein levels on GD20. ( H and I ) Fetal morphology and weight on GD20 across various groups. ( J and K ) Placental morphology and weight on GD20 in different groups. ( L ) Representative HE staining of kidney sections showing pathological changes. Scale bar: 100 μm. ( M and N ) IHC staining of CD31 in placental tissues. Scale bar: 100 μm. ( O – R ) Analysis of THBS4, ITG α2, and phosphorylated AKT expression using Western blotting in each group. n=6. ns: no significance, * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Light Microscopy, Injection, Staining, Immunohistochemistry, Expressing, Western Blot

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Proteomic profiling of PMSCs and PMSC-Exos. ( A ) Volcano plot showing differentially expressed proteins in PMSC-Exos compared to PMSCs. ( B ) GO analysis of upregulated proteins in terms of cellular component. ( C ) GO analysis of upregulated proteins in terms of biological processes. ( D ) KEGG pathway enrichment analysis of upregulated proteins. ( E ) Relative expression levels of angiogenesis-related proteins in PMSC-Exos versus PMSCs. ( F and G ) Western blotting analysis of THBS4 expression in PMSCs and PMSC-Exos. ( H ) THBS4 mRNA expression determined by RT-qPCR. ( I and J ) Western blotting analysis of THBS4 expression in normal and PE placenta. ( K and L ) IHC staining of THBS4 in placental tissues. Scale bar: 100 μm. ( M ) Predicted protein-protein interactions of THBS4. ( N and O ) Western blotting analysis of phosphorylated AKT in HUVECs under different treatment conditions. n=3. n=10 (H-K), ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemistry, Protein-Protein interactions

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: THBS4 mediates the pro-angiogenic and pro-migratory effects of PMSC-Exos on endothelial cells. ( A ) Lentiviral transfection of PMSCs with THBS4 shRNA or control vectors. Scale bar: 100 μm. ( B – D ) qRT-PCR and Western blotting analysis confirming THBS4 knockdown efficiency in PMSCs. ( E and F ) Western blotting analysis of THBS4 expression in exosomes derived from shNC- and shTHBS4-transfected PMSCs. ( G ) Confocal microscopy showing Dil-labeled exosome uptake by HUVECs. Scale bar: 100 μm. ( H and I ) Tube formation assays assessing angiogenesis of HUVECs under different treatments. Scale bar: 100 μm. ( J and K ) Transwell assays evaluating HUVEC migration under different treatments. Scale bar: 200 μm. ( L and M ) Western blot analysis of phosphorylated AKT levels in HUVECs after indicated treatments. n=3. *** P <0.001, **** P <0.0001.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Transfection, shRNA, Control, Quantitative RT-PCR, Western Blot, Knockdown, Expressing, Derivative Assay, Confocal Microscopy, Labeling, Migration

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Integrin α2 mediates THBS4-induced migration, angiogenesis, and AKT phosphorylation in HUVECs. ( A and B ) immunofluorescence co-localization of THBS4 and integrin α2. Scale bars: 100 μm and 10 μm. ( C and D ) Western blotting analysis of integrin α2 expression. ( E and F ) Representative images of tube formation assays in HUVECs after various treatments. Scale bar: 100 μm. ( G and H ) Representative images of the migration assay after different treatments in HUVECs. Scale bar: 200 μm. ( I and J ) Western blotting analysis of AKT phosphorylation expression after different treatments in HUVECs. ( K and L ) integrin α2 protein expression in HUVECs. ( M and N ) Images of tubular structures formed by HUVECs in each treatment group. (Scale bar: 100 μm). ( O and P ) Images of migrated HUVECs in each group. Scale bar: 200 μm. ( Q and R ) Western blotting analysis of the expression of AKT phosphorylation in each group. n=3. ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Migration, Phospho-proteomics, Immunofluorescence, Western Blot, Expressing

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: The inhibition of the PI3K/AKT pathway counteracts the angiogenic and migratory enhancement mediated by THBS4/integrin α2. ( A and B ) Representative tube formation images of HUVECs under indicated treatments. Scale bar: 100 μm. ( C and D ) Representative pictures of migrated HUVECs from transwell assays. Scale bar: 200 μm. ( E and F ) Western blotting analysis of the expression of AKT phosphorylation in each group. n=3. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Inhibition, Western Blot, Expressing, Phospho-proteomics

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Schematic illustration of the proposed mechanism. PMSC-derived exosomes (PMSC-Exos) enhance angiogenesis and tube formation of HUVECs and improve pregnancy outcomes in preeclamptic rats through the THBS4/integrin α2 (ITGα2)/PI3K/AKT signaling axis. The upward-pointing arrow denotes an increase.

Article Snippet: After blocking with 5% bovine serum albumin (BSA) for 1 hour, the cells were incubated overnight at 4°C with primary antibodies against THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse) and integrin α2 (ab181548, Abcam, USA, rabbit).

Techniques: Derivative Assay

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: The therapeutic effect of THBS4-rich-PMSC-Exos in preeclamptic rats. ( A ) Schematic of our animal experiments. ( B ) Detection of sperm in vaginal smears on gestational day 1 (GD1) under a light microscope. ( C ) Location of placental injection (indicated by arrow). ( D and E ) Systolic blood pressure (SBP) measurements and urinary protein levels on GD15. ( F and G ) Systolic blood pressure (SBP) measurements and urinary protein levels on GD20. ( H and I ) Fetal morphology and weight on GD20 across various groups. ( J and K ) Placental morphology and weight on GD20 in different groups. ( L ) Representative HE staining of kidney sections showing pathological changes. Scale bar: 100 μm. ( M and N ) IHC staining of CD31 in placental tissues. Scale bar: 100 μm. ( O – R ) Analysis of THBS4, ITG α2, and phosphorylated AKT expression using Western blotting in each group. n=6. ns: no significance, * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Light Microscopy, Injection, Staining, Immunohistochemistry, Expressing, Western Blot

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Proteomic profiling of PMSCs and PMSC-Exos. ( A ) Volcano plot showing differentially expressed proteins in PMSC-Exos compared to PMSCs. ( B ) GO analysis of upregulated proteins in terms of cellular component. ( C ) GO analysis of upregulated proteins in terms of biological processes. ( D ) KEGG pathway enrichment analysis of upregulated proteins. ( E ) Relative expression levels of angiogenesis-related proteins in PMSC-Exos versus PMSCs. ( F and G ) Western blotting analysis of THBS4 expression in PMSCs and PMSC-Exos. ( H ) THBS4 mRNA expression determined by RT-qPCR. ( I and J ) Western blotting analysis of THBS4 expression in normal and PE placenta. ( K and L ) IHC staining of THBS4 in placental tissues. Scale bar: 100 μm. ( M ) Predicted protein-protein interactions of THBS4. ( N and O ) Western blotting analysis of phosphorylated AKT in HUVECs under different treatment conditions. n=3. n=10 (H-K), ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunohistochemistry, Protein-Protein interactions

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: THBS4 mediates the pro-angiogenic and pro-migratory effects of PMSC-Exos on endothelial cells. ( A ) Lentiviral transfection of PMSCs with THBS4 shRNA or control vectors. Scale bar: 100 μm. ( B – D ) qRT-PCR and Western blotting analysis confirming THBS4 knockdown efficiency in PMSCs. ( E and F ) Western blotting analysis of THBS4 expression in exosomes derived from shNC- and shTHBS4-transfected PMSCs. ( G ) Confocal microscopy showing Dil-labeled exosome uptake by HUVECs. Scale bar: 100 μm. ( H and I ) Tube formation assays assessing angiogenesis of HUVECs under different treatments. Scale bar: 100 μm. ( J and K ) Transwell assays evaluating HUVEC migration under different treatments. Scale bar: 200 μm. ( L and M ) Western blot analysis of phosphorylated AKT levels in HUVECs after indicated treatments. n=3. *** P <0.001, **** P <0.0001.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Transfection, shRNA, Control, Quantitative RT-PCR, Western Blot, Knockdown, Expressing, Derivative Assay, Confocal Microscopy, Labeling, Migration

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Integrin α2 mediates THBS4-induced migration, angiogenesis, and AKT phosphorylation in HUVECs. ( A and B ) immunofluorescence co-localization of THBS4 and integrin α2. Scale bars: 100 μm and 10 μm. ( C and D ) Western blotting analysis of integrin α2 expression. ( E and F ) Representative images of tube formation assays in HUVECs after various treatments. Scale bar: 100 μm. ( G and H ) Representative images of the migration assay after different treatments in HUVECs. Scale bar: 200 μm. ( I and J ) Western blotting analysis of AKT phosphorylation expression after different treatments in HUVECs. ( K and L ) integrin α2 protein expression in HUVECs. ( M and N ) Images of tubular structures formed by HUVECs in each treatment group. (Scale bar: 100 μm). ( O and P ) Images of migrated HUVECs in each group. Scale bar: 200 μm. ( Q and R ) Western blotting analysis of the expression of AKT phosphorylation in each group. n=3. ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Migration, Phospho-proteomics, Immunofluorescence, Western Blot, Expressing

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: The inhibition of the PI3K/AKT pathway counteracts the angiogenic and migratory enhancement mediated by THBS4/integrin α2. ( A and B ) Representative tube formation images of HUVECs under indicated treatments. Scale bar: 100 μm. ( C and D ) Representative pictures of migrated HUVECs from transwell assays. Scale bar: 200 μm. ( E and F ) Western blotting analysis of the expression of AKT phosphorylation in each group. n=3. * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Inhibition, Western Blot, Expressing, Phospho-proteomics

Journal: International Journal of Nanomedicine

Article Title: Exosomal THBS4 Derived From Human Placental Mesenchymal Stem Cells Promotes the Migration and Angiogenesis of the Endothelial Cells in Preeclampsia by Activating the Integrin α2/PI3K/AKT Axis

doi: 10.2147/IJN.S572708

Figure Lengend Snippet: Schematic illustration of the proposed mechanism. PMSC-derived exosomes (PMSC-Exos) enhance angiogenesis and tube formation of HUVECs and improve pregnancy outcomes in preeclamptic rats through the THBS4/integrin α2 (ITGα2)/PI3K/AKT signaling axis. The upward-pointing arrow denotes an increase.

Article Snippet: The primary antibodies utilized were CD31 (ab182981, Abcam, USA, rabbit, 1:4000) and THBS4 (sc-28293, Santa Cruz Biotechnology, USA, mouse, 1:200).

Techniques: Derivative Assay

Journal: bioRxiv

Article Title: Regulatory T cells clonally expand and contribute to stromal cell function in fibrotic response to synthetic implants

doi: 10.64898/2026.01.05.697727

Figure Lengend Snippet: A) Scaled average expression (color) and percentage of expressing cells (dot size) of secreted ligands in each Treg cluster. B) Schematic of communication analysis from Tregs to fibroblasts and endothelial cells from the CD45-enriched scRNAseq data from Ruta et al. C) Communication score summarizing inferred signaling from Treg clusters to fibroblast and endothelial populations from (B). D) Activation of TFs in Treg to fibroblast network by fibroblast subcluster. E) Network plot illustrating inferred Treg to fibroblast signaling mediated by Sox family TFs with nodes representing sending clusters, ligands, receptors, TFs, and receiving clusters. Node size and edge thickness correspond to number of connections. F) Chord diagram showing connections from receptors and Sox family TFs for each fibroblast population. G) Mean expression of most highly expressed genes in Sox4 regulon in Lrrc15 + myofibroblast cluster, with matrisome associated genes indicated in blue. H) Schematic of fibroblast co-culture, performed with fibroblasts alone, or fibroblasts with Tregs from iLN or quadricep 6 weeks after PCL implant. I) Quantification via PCR of fibroblasts RNA after co-culture for Col1a2 , Col6a1 , and Thbs4 . J) Representative images of immunofluorescence staining of DAPI (white) and collagen I (green) in fibroblast coculture wells. Scale bar = 100µm. Statistical analysis was performed for technical replicates using one-way ANOVA with Tukey’s post hoc test. p < 0.05 (*), p < 0.01 (**), p < 0.001 (***).

Article Snippet: Murine TaqMan gene expression probes were used: Rer1 (Mm00471276_m1), Col1a2 (Mm00483888_m1), Col6a1 (Mm00487160_m1), and Thbs4 (Mm00449057_m1).

Techniques: Expressing, Activation Assay, Co-Culture Assay, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Thrombospondin-4 regulates apoptosis of vascular smooth muscle cells after artery transplanted into vein

doi: 10.1038/s41598-025-29180-2

Figure Lengend Snippet: High-throughput detection results after arterial grafting. ( A ) Heatmap of the top 100 upregulated differentially expressed genes in arterial grafts 14 days post-grafting. "C" represents the contralateral control group. “T” represents the experimental group 14 days after the surgery. ( B ) Heatmap of the top 150 differentially expressed proteins of arterial graft remodeling 30 days post-operation. "C" represents the contralateral control group. “T” represents the experimental group 30 days after the surgery. ( C ) THBS4 protein expression was detected via immunohistochemistry at 3, 7, 14, and 30 days post-operation . Neg represents the control group without adding the primary antibody. Red arrows indicate THBS4 protein expression in the cytoplasm and nucleus. Scale bars: 20 µm. ( D ) Comparison of the average optical density of immunohistochemical staining for THBS4 between groups. Data are expressed as mean ± standard deviation and measured by one-way ANOVA with Tukey’s post-hoc test for multiple comparisons (n = 4). * p < 0.0001 compared to 7 days post-operation.

Article Snippet: The rest were dewaxed with xylene, hydrated with ethanol, and incubated overnight at 4 °C with a 1:50 dilution of THBS4 primary antibody (SC-28293, Santa Cruz).

Techniques: High Throughput Screening Assay, Control, Expressing, Immunohistochemistry, Comparison, Immunohistochemical staining, Staining, Standard Deviation

Journal: Scientific Reports

Article Title: Thrombospondin-4 regulates apoptosis of vascular smooth muscle cells after artery transplanted into vein

doi: 10.1038/s41598-025-29180-2

Figure Lengend Snippet: Cell apoptosis and cycle arrest were induced in HASMCs by overexpressed THBS4. ( A ) Cell apoptosis analysis of HASMCs after THBS4 overexpression for 48 h was detected via flow cytometry analysis using Annexin V-APC and 7-AAD double staining. HASMCs are presented in a scatter plot where the upper left quadrant (Q1) identifies naked nucleated cells (7-AAD + /APC - ), the upper right quadrant (Q2) identifies late apoptotic cells (7-AAD + /APC + ), the lower left quadrant (Q3) identifies live cells (7-AAD − /APC − ), and the lower right quadrant (Q4) identifies early apoptotic cells (7-AAD - /APC + ). ( B ) Quantification of apoptotic cells by THBS4 overexpression. The percentage of apoptotic cells was calculated. Apoptosis rate = the upper right quadrant (Q2) + the lower right quadrant (Q4). Data are expressed as mean ± standard deviation and analyzed by unpaired T-test. n = 3 experiments, ** p < 0.01. ( C ) Cell apoptosis analysis of HASMCs after THBS4 silencing for 48 h was detected via flow cytometry analysis using Annexin V-APC and 7-AAD double staining. ( D ) Quantification of apoptotic cells by THBS4 silencing. Data are expressed as mean ± standard deviation and analyzed by unpaired T-test. n = 3 experiments, ns = non-significant. ( E ) Changes in the cell cycle of HASMCs owing to overexpressed THBS4 for 48 h. ( F ) Percentages of different cell cycles in HASMCs after THBS4 was overexpressed. Percentage of cells in the S phase increased significantly. Data are expressed as mean ± standard deviation and analyzed by unpaired T-test. n = 3 experiments, ** p < 0.01. ( G ) Changes in the cell cycles of HASMCs owing to silenced THBS4 for 48 h. (H) Percentages of different cell cycles in HASMCs after silencing THBS4. Data are expressed as mean ± standard deviation and analyzed by unpaired T-test. n = 3 experiments, ns = non-significant.

Article Snippet: The rest were dewaxed with xylene, hydrated with ethanol, and incubated overnight at 4 °C with a 1:50 dilution of THBS4 primary antibody (SC-28293, Santa Cruz).

Techniques: Over Expression, Flow Cytometry, Double Staining, Standard Deviation

Journal: Scientific Reports

Article Title: Thrombospondin-4 regulates apoptosis of vascular smooth muscle cells after artery transplanted into vein

doi: 10.1038/s41598-025-29180-2

Figure Lengend Snippet: THBS4 expression and apoptosis in VSMCs under different intensities of mechanical stretch. ( A ) THBS4 gene expression at stretch deformation 0,5,10 and 15%. Data are expressed as mean ± standard deviation and measured by One-way ANOVA with Tukey’s post-hoc test for multiple comparisons. n = 3 experiments, p < 0.05, significant difference, #compared to stretch deformation 5%. ( B ) Quantification of THBS4 protein in HASMCs at stretch deformation 0, 5, 10 and 15%. Data are expressed as mean ± standard deviation and measured by One-way ANOVA with Tukey’s post-hoc test for multiple comparisons. n = 3 experiments, p < 0.05, significant difference, ⊙compared to stretch deformation 15%. ( C ) The levels of THBS4 in HASMCs were detected by western blotting undert stretch deformation of 0, 5, 10 and 15%. The original blots are presented in Supplementary Fig. . ( D ) Apoptosis analysis of HASMCs at stretch deformation 0,5,10 and 15% was detected via flow cytometry analysis using Annexin V-APC and 7-AAD double staining. ( E ) Quantification of apoptotic cells by different stretch deformation. Data are expressed as mean ± standard deviation and analyzed by One-way ANOVA with Tukey’s post-hoc test for multiple comparisons, n = 3 experiments, p < 0.05, significant difference, *compared to stretch deformation 0%, #compared to stretch deformation 5%.

Article Snippet: The rest were dewaxed with xylene, hydrated with ethanol, and incubated overnight at 4 °C with a 1:50 dilution of THBS4 primary antibody (SC-28293, Santa Cruz).

Techniques: Expressing, Gene Expression, Standard Deviation, Western Blot, Flow Cytometry, Double Staining

Journal: Scientific Reports

Article Title: Thrombospondin-4 regulates apoptosis of vascular smooth muscle cells after artery transplanted into vein

doi: 10.1038/s41598-025-29180-2

Figure Lengend Snippet: THBS4 regulates VSMCs apoptosis, interacting with the focal adhesion pathway. ( A ) IPG analysis of the perturbation of the focal adhesion pathway. Blue indicates downregulated genes; Red indicates upregulated genes. Pathway analysis was performed using KEGG database , . ( B ) The molecular mechanisms related to the apoptotic process may be regulated in the focal adhesion pathways. Blue indicates downregulated genes; Red indicates upregulated genes. ( C ) p-PAK1 expression after transfection with THBS4 for 72 h. p-PAK1 expression was upregulated after THBS4 was overexpressed on the left. On the right, p-PAK1 expression was inhibited after small RNA interfered with THBS4. Data are expressed as mean ± standard deviation and analzsed by paired T-test. n = 6 experiments, * p < 0.05. The original blots are presented in Supplementary Figs. and .

Article Snippet: The rest were dewaxed with xylene, hydrated with ethanol, and incubated overnight at 4 °C with a 1:50 dilution of THBS4 primary antibody (SC-28293, Santa Cruz).

Techniques: Expressing, Transfection, Standard Deviation