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gene exp thbs4 mm00449057 m1  (Thermo Fisher)
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Thermo Fisher gene exp thbs4 mm00449057 m1
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 <t>Thbs4</t> . 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 (***).
Gene Exp Thbs4 Mm00449057 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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si thbs4 tgf β1  (MedChemExpress)
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MedChemExpress si thbs4 tgf β1
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 <t>Thbs4</t> . 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 (***).
Si Thbs4 Tgf β1, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/si thbs4 tgf β1/product/MedChemExpress
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gene exp thbs4 hs00170261 m1  (Thermo Fisher)
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Thermo Fisher gene exp thbs4 hs00170261 m1
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 <t>Thbs4</t> . 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 (***).
Gene Exp Thbs4 Hs00170261 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/gene exp thbs4 hs00170261 m1/product/Thermo Fisher
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primepcr ddpcr gene expression probe assay fam thbs4  (Bio-Rad)
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Bio-Rad primepcr ddpcr gene expression probe assay fam thbs4
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 <t>Thbs4</t> . 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 (***).
Primepcr Ddpcr Gene Expression Probe Assay Fam Thbs4, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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murine thbs4  (Addgene inc)
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Addgene inc murine thbs4
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 <t>Thbs4</t> . 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 (***).
Murine Thbs4, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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plasmid containing murine thbs4 paav.cmv.sv40.thbs4ha.sv40(polya)  (Addgene inc)
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Addgene inc plasmid containing murine thbs4 paav.cmv.sv40.thbs4ha.sv40(polya)
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 <t>Thbs4</t> . 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 (***).
Plasmid Containing Murine Thbs4 Paav.Cmv.Sv40.Thbs4ha.Sv40(Polya), supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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antibodies against thbs4  (Danaher Inc)
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Danaher Inc antibodies against thbs4
Circulating level of <t>THBS4</t> in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05
Antibodies Against Thbs4, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human thbs4 elisa kit  (Cusabio)
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Cusabio human thbs4 elisa kit
Circulating level of <t>THBS4</t> in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05
Human Thbs4 Elisa Kit, supplied by Cusabio, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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recombinant human insulin-like growth factor-1  (MedChemExpress)
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MedChemExpress recombinant human insulin-like growth factor-1
Circulating level of <t>THBS4</t> in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05
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Image Search Results


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 (***).

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

Circulating level of THBS4 in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Circulating level of THBS4 in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques:

Expression of THBS4 in PAH rats induced by MCT plus aorto-caval shunt. (A) THBS4 expression in GSE149899. qRT-PCR (B) and Western blot (C) showed the time course of THBS4 expression in PAH rats induced by MCT plus aorto-caval shunt. (D) Double immunofluorescence staining of THBS4 with α-SMA or VWF. Scare bars: 50 μm. CON: the control group; SHAM: the sham group; MS: PAH rats induced by MCT plus aorto-caval shunt (MS) from 1 week (1w) to 4 weeks (4w). The resultant data are represented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Expression of THBS4 in PAH rats induced by MCT plus aorto-caval shunt. (A) THBS4 expression in GSE149899. qRT-PCR (B) and Western blot (C) showed the time course of THBS4 expression in PAH rats induced by MCT plus aorto-caval shunt. (D) Double immunofluorescence staining of THBS4 with α-SMA or VWF. Scare bars: 50 μm. CON: the control group; SHAM: the sham group; MS: PAH rats induced by MCT plus aorto-caval shunt (MS) from 1 week (1w) to 4 weeks (4w). The resultant data are represented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Immunofluorescence Staining, Control

qRT-PCR (A) and Western blot (B) showed the transfection efficiency of THBS4 in PASMCs. (C) The influence of THBS4 on the transformation of phenotype of PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; *** P < 0.001; # P < 0.05; ## P < 0.01; ### P < 0.001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: qRT-PCR (A) and Western blot (B) showed the transfection efficiency of THBS4 in PASMCs. (C) The influence of THBS4 on the transformation of phenotype of PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; *** P < 0.001; # P < 0.05; ## P < 0.01; ### P < 0.001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Transformation Assay

Effect of THBS4 on the proliferation, apoptosis, and migration of PASMCs. (A) Effect of THBS4 on proliferation of PASMCs, as determined by the EdU assay. Scare bars, 100 μm. (B) Effect of THBS4 on apoptosis of PASMCs, as determined by the Annexin-V flow cytometry. (C) Effect of THBS4 on migration of PASMCs, as determined by the Wound healing assay. Scare bars, 200 μm. (D) Effect of THBS4 on migration of PASMCs, as determined by the Transwell assay. Scare bars, 100 μm. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; ** P < 0.01; **** P < 0.0001; # P < 0.05; ## P < 0.01

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 on the proliferation, apoptosis, and migration of PASMCs. (A) Effect of THBS4 on proliferation of PASMCs, as determined by the EdU assay. Scare bars, 100 μm. (B) Effect of THBS4 on apoptosis of PASMCs, as determined by the Annexin-V flow cytometry. (C) Effect of THBS4 on migration of PASMCs, as determined by the Wound healing assay. Scare bars, 200 μm. (D) Effect of THBS4 on migration of PASMCs, as determined by the Transwell assay. Scare bars, 100 μm. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; ** P < 0.01; **** P < 0.0001; # P < 0.05; ## P < 0.01

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Migration, EdU Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay

Phosphorylation level of PI3K/AKT in PASMCs after inhibition and overexpression of THBS4 in PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs. si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.5; # P < 0.5

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Phosphorylation level of PI3K/AKT in PASMCs after inhibition and overexpression of THBS4 in PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs. si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.5; # P < 0.5

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Inhibition, Over Expression

Effect of THBS4 suppression on pulmonary vascular remodeling in PAH rats induced by MCT plus aorto-caval shunt. (A) The AAV transfection efficiency was determined by detecting green fluorescence protein which is encoded by AAV in lung tissues. Scare bars, 100 μm. (B) THBS4 suppression efficiency determined by immunofluorescence staining of THBS4. Scare bars, 50 μm. (C) Hematoxylin and Eosin staining of lung tissues. Scare bars,100 μm. (D) Immunofluorescence staining of α-SMA in lung tissues. Scare bars,100 μm. The percentage of non-muscularized (E) , partially-muscularized (F), and fully-muscularized pulmonary arterioles (G) . (H) The percentage of wall thickness. (I) The percentage of wall area. (G) Cross-sectional area of the right ventricular cardiomyocyte. Scare bars,100 μm. The resultant data are represented as mean ± SD. CON: control group; SHAM: sham group; MS-AAV-CON: administration of control AAV + MCT plus aorto-caval shunt induced PAH rats; MS-AAV-siTHBS4: administration of THBS4 suppression AAV + MCT plus aorto-caval shunt induced PAH rats. PA: pulmonary arteriole. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 suppression on pulmonary vascular remodeling in PAH rats induced by MCT plus aorto-caval shunt. (A) The AAV transfection efficiency was determined by detecting green fluorescence protein which is encoded by AAV in lung tissues. Scare bars, 100 μm. (B) THBS4 suppression efficiency determined by immunofluorescence staining of THBS4. Scare bars, 50 μm. (C) Hematoxylin and Eosin staining of lung tissues. Scare bars,100 μm. (D) Immunofluorescence staining of α-SMA in lung tissues. Scare bars,100 μm. The percentage of non-muscularized (E) , partially-muscularized (F), and fully-muscularized pulmonary arterioles (G) . (H) The percentage of wall thickness. (I) The percentage of wall area. (G) Cross-sectional area of the right ventricular cardiomyocyte. Scare bars,100 μm. The resultant data are represented as mean ± SD. CON: control group; SHAM: sham group; MS-AAV-CON: administration of control AAV + MCT plus aorto-caval shunt induced PAH rats; MS-AAV-siTHBS4: administration of THBS4 suppression AAV + MCT plus aorto-caval shunt induced PAH rats. PA: pulmonary arteriole. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: The PVDF membrane was blocked for nonspecific binding with blocking buffer and incubated with primary antibodies against THBS4 (Abcam, USA, 1:1000), GAPDH (Proteintech, China, 1:20000), α-SMA (Proteintech, China, 1:20000), MYH11 (SANTA CRUZ, America, 1:200), PCNA (SANTA CRUZ, America, 1:200), N-cad (SANTA CRUZ, America, 1:200), BAX (Abmart, China, 1:1000), AKT (CST, America,1:1000), PI3K (CST, America, 1:1000), p-AKT (Abmart, China, 1:1000), and p-PI3K (Abmart, China, 1:1000) overnight at 4 ℃.

Techniques: Transfection, Fluorescence, Immunofluorescence, Staining, Control

Circulating level of THBS4 in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Circulating level of THBS4 in patients with PAH-CHD. The resultant data are represented as mean ± SD. * P < 0.05

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques:

Expression of THBS4 in PAH rats induced by MCT plus aorto-caval shunt. (A) THBS4 expression in GSE149899. qRT-PCR (B) and Western blot (C) showed the time course of THBS4 expression in PAH rats induced by MCT plus aorto-caval shunt. (D) Double immunofluorescence staining of THBS4 with α-SMA or VWF. Scare bars: 50 μm. CON: the control group; SHAM: the sham group; MS: PAH rats induced by MCT plus aorto-caval shunt (MS) from 1 week (1w) to 4 weeks (4w). The resultant data are represented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Expression of THBS4 in PAH rats induced by MCT plus aorto-caval shunt. (A) THBS4 expression in GSE149899. qRT-PCR (B) and Western blot (C) showed the time course of THBS4 expression in PAH rats induced by MCT plus aorto-caval shunt. (D) Double immunofluorescence staining of THBS4 with α-SMA or VWF. Scare bars: 50 μm. CON: the control group; SHAM: the sham group; MS: PAH rats induced by MCT plus aorto-caval shunt (MS) from 1 week (1w) to 4 weeks (4w). The resultant data are represented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Double Immunofluorescence Staining, Control

qRT-PCR (A) and Western blot (B) showed the transfection efficiency of THBS4 in PASMCs. (C) The influence of THBS4 on the transformation of phenotype of PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; *** P < 0.001; # P < 0.05; ## P < 0.01; ### P < 0.001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: qRT-PCR (A) and Western blot (B) showed the transfection efficiency of THBS4 in PASMCs. (C) The influence of THBS4 on the transformation of phenotype of PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; *** P < 0.001; # P < 0.05; ## P < 0.01; ### P < 0.001

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Western Blot, Transfection, Transformation Assay

Effect of THBS4 on the proliferation, apoptosis, and migration of PASMCs. (A) Effect of THBS4 on proliferation of PASMCs, as determined by the EdU assay. Scare bars, 100 μm. (B) Effect of THBS4 on apoptosis of PASMCs, as determined by the Annexin-V flow cytometry. (C) Effect of THBS4 on migration of PASMCs, as determined by the Wound healing assay. Scare bars, 200 μm. (D) Effect of THBS4 on migration of PASMCs, as determined by the Transwell assay. Scare bars, 100 μm. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; ** P < 0.01; **** P < 0.0001; # P < 0.05; ## P < 0.01

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 on the proliferation, apoptosis, and migration of PASMCs. (A) Effect of THBS4 on proliferation of PASMCs, as determined by the EdU assay. Scare bars, 100 μm. (B) Effect of THBS4 on apoptosis of PASMCs, as determined by the Annexin-V flow cytometry. (C) Effect of THBS4 on migration of PASMCs, as determined by the Wound healing assay. Scare bars, 200 μm. (D) Effect of THBS4 on migration of PASMCs, as determined by the Transwell assay. Scare bars, 100 μm. The resultant data are represented as mean ± SD. *: si-THBS4 vs.si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.05; ** P < 0.01; **** P < 0.0001; # P < 0.05; ## P < 0.01

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques: Migration, EdU Assay, Flow Cytometry, Wound Healing Assay, Transwell Assay

Phosphorylation level of PI3K/AKT in PASMCs after inhibition and overexpression of THBS4 in PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs. si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.5; # P < 0.5

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Phosphorylation level of PI3K/AKT in PASMCs after inhibition and overexpression of THBS4 in PASMCs. The resultant data are represented as mean ± SD. *: si-THBS4 vs. si-CON; #: pEX3-THBS4 vs. pEX3-CON. * P < 0.5; # P < 0.5

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques: Phospho-proteomics, Inhibition, Over Expression

Effect of THBS4 suppression on pulmonary vascular remodeling in PAH rats induced by MCT plus aorto-caval shunt. (A) The AAV transfection efficiency was determined by detecting green fluorescence protein which is encoded by AAV in lung tissues. Scare bars, 100 μm. (B) THBS4 suppression efficiency determined by immunofluorescence staining of THBS4. Scare bars, 50 μm. (C) Hematoxylin and Eosin staining of lung tissues. Scare bars,100 μm. (D) Immunofluorescence staining of α-SMA in lung tissues. Scare bars,100 μm. The percentage of non-muscularized (E) , partially-muscularized (F), and fully-muscularized pulmonary arterioles (G) . (H) The percentage of wall thickness. (I) The percentage of wall area. (G) Cross-sectional area of the right ventricular cardiomyocyte. Scare bars,100 μm. The resultant data are represented as mean ± SD. CON: control group; SHAM: sham group; MS-AAV-CON: administration of control AAV + MCT plus aorto-caval shunt induced PAH rats; MS-AAV-siTHBS4: administration of THBS4 suppression AAV + MCT plus aorto-caval shunt induced PAH rats. PA: pulmonary arteriole. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Journal: Respiratory Research

Article Title: The role and mechanism of thrombospondin-4 in pulmonary arterial hypertension associated with congenital heart disease

doi: 10.1186/s12931-024-02932-w

Figure Lengend Snippet: Effect of THBS4 suppression on pulmonary vascular remodeling in PAH rats induced by MCT plus aorto-caval shunt. (A) The AAV transfection efficiency was determined by detecting green fluorescence protein which is encoded by AAV in lung tissues. Scare bars, 100 μm. (B) THBS4 suppression efficiency determined by immunofluorescence staining of THBS4. Scare bars, 50 μm. (C) Hematoxylin and Eosin staining of lung tissues. Scare bars,100 μm. (D) Immunofluorescence staining of α-SMA in lung tissues. Scare bars,100 μm. The percentage of non-muscularized (E) , partially-muscularized (F), and fully-muscularized pulmonary arterioles (G) . (H) The percentage of wall thickness. (I) The percentage of wall area. (G) Cross-sectional area of the right ventricular cardiomyocyte. Scare bars,100 μm. The resultant data are represented as mean ± SD. CON: control group; SHAM: sham group; MS-AAV-CON: administration of control AAV + MCT plus aorto-caval shunt induced PAH rats; MS-AAV-siTHBS4: administration of THBS4 suppression AAV + MCT plus aorto-caval shunt induced PAH rats. PA: pulmonary arteriole. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: A human THBS4 ELISA kit (Cusabio, China) was used to measure the circulating level of THBS4 according to the manufacturer’s instructions.

Techniques: Transfection, Fluorescence, Immunofluorescence, Staining, Control