human thbs4 elisa kit  (Cusabio)

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:

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

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

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

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

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

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Heatmap of serum proteomics of Bambi fl/fl and Bambi HSA-Cre mice. (B) Principal coordinate analysis (PCoA) of serum proteomics of Bambi fl/fl and Bambi HSA-Cre mice. (C) GO analysis (Biological Process) of differentially upregulated protein biomarkers of Bambi fl/fl and Bambi HSA-Cre mice. (D) qPCR analysis of Thbs4 expression in different tissues of C57BL/6J mice. (E) qPCR analysis of Thbs4 expression in TA and iWAT of Bambi fl/fl and Bambi HSA-Cre mice (n=5 mice/group). (F) Representative immunoblots of Thbs4 protein levels in the supernatant of ex vivo cultured TA muscle for 72-hr of Bambi fl/fl and Bambi HSA-Cre mice. (G) Representative images of H&E staining. and IHC of Thbs4, PGC1α and Ucp1 in iBAT sections from the RT and cold challenge groups. (H) Representative images of H&E staining. and IHC of Thbs4, PGC1α and Ucp1 in eWAT sections at the condition of R.T. or cold challenge for 7 days. ND, not detected, N.S., not significant, * p < 0.05, and ** p < 0.01 by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Expressing, Western Blot, Ex Vivo, Cell Culture, Staining

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Venn diagram indicating overlapping genes between the TA muscle transcriptome and matched serum proteomics between Bambi fl/fl and Bambi HSA-Cre mice. (B) Venn diagram and heatmap showing overlapping genes in TA muscle of Bambi fl/fl and Bambi HSA-Cre mice against the secretory protein library. (C) Thbs4 serum levels of Bambi fl/fl and Bambi HSA-Cre mice (n=5 mice/group). (D) Representative immunoblots of Thbs4 expression in iWAT and serum. (E) IHC staining of Thbs4 in the iWAT between Bambi fl/fl and Bambi HSA-Cre mice. (F) Diagram depicting the timeline of cold challenge for C57BL/6J mice. (G) Representative immunoblots showing the levels of Thbs4 in serum and iWAT from C57BL/6J mice with cold challenge. (H) qPCR analysis of Thbs4 expression levels in the TA muscle of C57BL/6J mice. Different letters indicate significant differences between two groups. (I) Representative images of H&E, Thbs4, PGC1α, and Ucp1 IHC in iWAT sections from C57BL/6J mice at the condition of R.T. or cold challenge for 7 days. * p < 0.05, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Western Blot, Expressing, Immunohistochemistry

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) The rate of body weight gain in the sedentary and 30-day aerobic training of treadmill running groups (n=4 mice/group of two independent repeat). (B) Serum Thbs4 levels in the sedentary and treadmill running groups. (C-D) GTT and ITT for the sedentary and treadmill running groups. (E) Representative images of H&E staining and IHC staining of Thbs4, PGC1α and Ucp1 antibodies in the iWAT of mice in the sedentary and treadmill running groups. (F) qPCR analysis of thermogenesis-related gene expression in the iWAT of mice in the sedentary and treadmill running groups. (G) Representative immunoblots of Thbs4 (TA muscle) and Ucp1 (iWAT) in the sedentary and treadmill running groups. (H) Representative images of IHC staining of Glut4 in the iWAT of mice in the sedentary and treadmill running groups. (I) Representative immunoblots of Thbs4 protein levels in serum in the sedentary and treadmill running groups. (J) Serum Thbs4 protein levels in the sedentary and treadmill running groups. (K) The percentage of lean mass and fat mass in the sedentary and treadmill running groups. N.S., not significant, * p < 0.05, ** p < 0.01 and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Staining, Immunohistochemistry, Gene Expression, Western Blot

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Distribution of ATAC-seq peaks in the whole genome across promoters, introns, exon distal intergenic regions and UTRs. (B) Motif enrichment analysis of the peaks aligned in the chromatin opening region. (C) Venn diagrams of overlapping DEGs between RNA-seq (between Bambi fl/fl and Bambi HSA-Cre mice) and ATAC-seq (peaks associated unique genes). (D) Heatmap of 681 overlapping DEGs between Bambi fl/fl and Bambi HSA-Cre mice. (E) KEGG pathway enrichment analysis of upregulated genes in TA muscle from Bambi fl/fl and Bambi HSA- Cre mice 30 days post-CTX-induced injury. (F) UCSC genome browser traces of ATAC-seq peaks at the promoter regions of the Pax7 , Myod1 , Myog , Myf5 , and Thbs4 loci. Identification of a conserved MyoD/MyoG binding site in the proximal promoter region of Thbs4. (G) Representative images of Pax7 (Red), Thbs4 (Green), and MyoD (Purple) immunostaining of the SCs (day 0) or cultured for 72-hr (day 3) in single myofiber (n>100 clusters from n=3 mice/group). (H) qPCR analysis of Myod1 , Myog and Thbs4 expression in TA muscle post CTX-induced injury on T0, T3, T5, T7, T9, T12 and T15 (n=6 mice). (I) Representative immunostaining images of Thbs4 (Green) and myogenin (Red) in C2C12 myoblasts at the proliferation (day 0) and differentiation stages (day 5). (J) Representative immunoblots of Thbs4 protein levels in C2C12 myoblast lysate and culture medium with either Ctrl or Thbs4-expressing adenovirus infection with/without BFA treatment at 0, 1, and 2 μg/ml. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: RNA Sequencing, Binding Assay, Immunostaining, Cell Culture, Expressing, Western Blot, Infection

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Experimental scheme of intramuscular injection of AAV-Ctrl and AAV-Thbs4 groups (n=3 mice/group). (B) Representative immunoblots of Thbs4 protein in TA muscle and serum in the AAV-Ctrl and AAV-Thbs4 groups. (C) Representative images of IHC staining of Thbs4 and Ucp1 in the iWAT of mice with either AAV-Ctrl or AAV-Thbs4 intramuscular injection. (D) qPCR analysis of Prdm16 , Ucp1 , Ppargc1α , Cox7a1 , Cox8b mRNA expression levels in iWAT in the AAV-Ctrl and AAV-Thbs4 groups. (E) Representative images of mice on HFD with intramuscular injection of either AAV-Ctrl or AAV-Thbs4 (n=5 mice/group). (F) Daily food intake for mice in the AAV-Ctrl and AAV-Thbs4 groups. (G) Body weight was measured weekly during the HFD feeding. (H-I) Glucose tolerance test (GTT) and AUC. (J) Serum levels of TG, TC, FFA, LDL-c and HDL-c between the AAV-Ctrl and AAV-Thbs4 groups. (K) qPCR analysis of thermogenesis- and beige fat-related gene expression in iWAT in the AAV-Ctrl and AAV-Thbs4 groups. N.S., not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Injection, Western Blot, Immunohistochemistry, Expressing, Gene Expression

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Body weight of the AAV-Ctrl and AAV-Thbs4 groups (n=3 mice/group). (B) The rectal temperature of the AAV-Ctrl and AAV-Thbs4 groups. (C) The tissue weight of TA muscle and iWAT of the AAV-Ctrl and AAV-Thbs4 groups. (D) IHC of Thbs4 and Ucp1 in the section of iBAT, Heart, Liver and Pancreas of C57BL6/J mice of the AAV-Ctrl or AAV-Thbs4 groups. N.S., not significant, and ** p < 0.01, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques:

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Representative images of TA muscle of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding (n=5 mice/group). (B) Representative images of iWAT of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (C) Tissue weight of TA, iBAT, iWAT, eWAT and liver of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (D) ITT for AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (E) AUC of ITT of AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (F) Serum levels of AST and ALT of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (G) Representative images of H&E staining of iBAT sections of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (H) Representative images of H&E staining of iWAT sections, whole section images (left) and snap images (right) of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (I) Representative images of eWAT (left) and H&E staining (right) of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (J) Frequency distribution of the size of lipid droplets in eWAT of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (K) Representative images of liver (left) and H&E staining (right) of AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. (L) Representative of immunoblots of Ucp1 and Tubulin of iWAT and Thbs4-HA of serum of the AAV-Ctrl and AAV-Thbs4 groups under HFD feeding. N.S., not significant, * p < 0.05, ** p < 0.01 and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Staining, Western Blot

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Average VO 2 (left) was monitored over 48-hr period and the AUC of VO 2 (right) curve for WT and Thbs4- KO mice at R.T. (n=4 mice/group of two independent repeats). (B) Average VCO 2 (left) was monitored over a 48-hr period and the AUC of VCO 2 (right) curve for WT and Thbs4- KO mice at R.T. (n=4 mice/group of two independent repeats). (C) qPCR analysis of Thbs4 expression in TA and Sol muscle of WT and Thbs4- KO mice (n=6 mice/group). (D) qPCR analysis of thermogenesis-related gene expression in iBAT of WT and Thbs4- KO mice (n=6 mice/group). (E) qPCR analysis of thermogenesis-related gene expression in iWAT of WT and Thbs4- KO mice (n=6 mice/group). (F) Seahorse flux analysis of OCR of undifferentiated SVFs isolated from iWAT of WT and Thbs4- KO mice. (G) Seahorse flux analysis of OCR of differentiated primary white adipocyte of WT and Thbs4- KO mice. (H) Statistical analysis of OCR at the stage of basal respiration, maximal respiration, proton leaky, ATP production and spare respiration capacity. (I) qPCR analysis of thermogenesis-related gene expression of differentiated primary white adipocyte of WT and Thbs4- KO mice. (J) Representative image of BODIPY (Green) staining of differentiated primary white adipocyte. (K) Representative immunoblots of Ucp1, PGC1α, Glut4, p-HSL (Ser565), t-HSL and Mito-complex. (L) Representative immunoblots of Ucp1 and Tubulin of differentiated primary adipocyte from WT, Thbs4 -KO and aged C57BL6/J mice. N.S., not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Expressing, Gene Expression, Isolation, Staining, Western Blot

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Diagram depicting the timeline of the HFD feeding plan for WT and Thbs4- KO mice (n=8 mice/group). (B) Body weight was measured weekly during HFD feeding. (C) Daily food intake for mice on HFD feeding over 25 weeks. (D) Serum levels of TG, TC, FFA, LDL-c and HDL-c (n=8 mice/group). (E-F) Glucose tolerance test (GTT) and insulin tolerance test (ITT). (G) Representative images of iBAT and H.&E. staining. (H) Representative images of liver and H.&E. staining. (I) Representative images of iWAT, H.&E. and F4/80 IHC staining. (J) Representative images of eWAT, H.&E. and F4/80 IHC staining. N.S., not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Staining, Immunohistochemistry

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) GTT and initial fasting glucose level of WT and Thbs4 -KO mice under HFD feeding (n=7-8 mice/group). (B) ITT of WT and Thbs4 -KO mice under HFD feeding (n=7-8 mice/group). (C) The rectal temperature of WT and Thbs4 -KO mice under HFD feeding before and after 16-hr fasting (n=7-8 mice/group). (D) Representative images of tissues (TA, Sol, EDL and Gas muscle) of WT and Thbs4 -KO mice under HFD feeding (n=7-8 mice/group). (E) The tissue weight of TA, EDL, Sol, Gas, iBAT, iWAT, eWAT and liver of WT and Thbs4 -KO mice under HFD feeding (n=7-8 mice/group). (F) qPCR analysis of lipid metabolism-related gene expression in liver of WT and Thbs4 -KO mice under HFD feeding (n=7-8 mice/group). N.S., not significant, * p < 0.05, and *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Gene Expression

Journal: bioRxiv

Article Title: A muscle hypertrophy-derived myokine reprograms the stromal vascular fractions differentiation towards thermogenic adipocytes in subcutaneous adipose tissue

doi: 10.1101/2023.09.20.558627

Figure Lengend Snippet: (A) Heatmap of the Thbs4 expression profile in TA, EDL, Soleus and Gas muscle of C57BL6/J mice at different ages (2wk, 8wk, 30wk, 60wk and 80wk) (n=3 mice/group). (B) qPCR analysis of Thbs4 expression in TA, EDL, Sol and Gas muscle of C57BL6/J mice between adult (8wk) and aged (80wk) (n=4). (C) Representative immunoblots of Thbs4 protein level in TA muscle of C57BL6/J mice between adult (8wk) and aged (80wk) (n=8). *** p < 0.001, by two-sided Student’s t test. Data represent the mean ± standard error of the mean.

Article Snippet: Serum Thbs4 levels were measured by using the Thbs4 ELISA kit (EL6512, Cusabio) by following the manufacture’s instruction.

Techniques: Expressing, Western Blot

Journal: Advanced Science

Article Title: Single‐Cell RNA Sequencing and Spatial Transcriptomics Reveal Pathogenesis of Meningeal Lymphatic Dysfunction after Experimental Subarachnoid Hemorrhage

doi: 10.1002/advs.202301428

Figure Lengend Snippet: Intercellular communication networks in and around mLVs. A) Spatial visualization of immune cell infiltration based on mLECs distribution. B) Outgoing and incoming signal patterns among all meningeal cells. C) THBS signaling pathway network among all cell types. D) Spatial visualization of THBS1‐CD47 L‐R pair distribution by stLearn. E) Volcano plots showing that THBS1 is one of the upregulated DEGs in the SAH group compared to the NPH group identified by mass spectrometry. F) Quantification of THBS1, THBS2, and THBS4 expression in CSF samples from SAH patients compared to those from NPH patients by ELISA assay, *** p < 0.001 versus NPH by paired two‐tailed Student's t ‐test. G) Representative confocal images of mLVs region of the negative control (NC) group and recombinant THBS1 protein treatment (rTHBS1) group. An enlarged view of TS and SSS is shown on the right in each group. Lyve1 (blue), Beads (green), and CD31 (red). Scale bar: 1000 µm. H) Flow cytometric analysis of mLECs percentage in NC group and rTHBS1 group, n = 4 per group, *** p < 0.001 versus NC by paired two‐tailed Student's t ‐test. I) Representative images of beads accumulation in dCLNs of the NC group and rTHBS1 group. Scale bar: 200 µm.

Article Snippet: The concentrations of human CSF THBS1 (EK0899, Boster Biotech, Wuhan), THBS2 (EK0642, Boster Biotech, Wuhan), THBS4 (CSB‐EL023490HU, Cusabio Biotech, Wuhan), and S100A6 (CSB‐EL020634RA, Cusabio Biotech, Wuhan) were determined using ELISA kits.

Techniques: Mass Spectrometry, Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Negative Control, Recombinant

Journal: Clinical and Translational Medicine

Article Title: Intestinal fibrosis in aganglionic segment of Hirschsprung's disease revealed by single‐cell RNA sequencing

doi: 10.1002/ctm2.1193

Figure Lengend Snippet: Source of extracellular matrix (ECM) in the intestine. (A) Heatmap of area under the curve (AUC) scores of collagens, glycoproteins, proteoglycans and all ECM gene set in merge datasets. The colour bars match the cell‐type group colours. (B) Heatmap of HSCR‐related ECM genes among different segments in merge dataset. The colour bars match the cell‐type group colours. (C) FeaturePlot of MFAP5 and THBS4 among different segments. (D) Violin plot of expression of MFAP5 among different samples in smooth muscle, myofibroblast and fibroblast. (E) Violin plot of expression of THBS4 among different samples in smooth muscle, myofibroblast and fibroblast. (F) Representative images of immunohistochemistry (IHC) for MFAP5 protein in the enteric ganglia and smooth muscle layers of the normal and aganglionic sections (i). Real‐time polymerase chain reaction (RT‐qPCR) validation of the MFAP5 gene in the normal and aganglionic segments per subject, p ‐value was calculated from Wilcoxon matched‐pairs signed‐rank test (ii). (G) Representative images of IHC for THBS4 protein in the enteric ganglia and smooth muscle layers of the normal and aganglionic sections (i). RT‐qPCR validation of the THBS4 gene in the normal and aganglionic segments per subject, p ‐value was calculated from Wilcoxon matched‐pairs signed‐rank test (ii). UMAP, uniform manifold approximation and projection.

Article Snippet: To analyse the expression location of target DEGs in HSCR samples, IHC for MFAP5 (Abcam, ab203828, Rabbit monoclonal immunoglobulin G (IgG), 1:5000 dilution), THBS4 (Abcam, ab263898, Rabbit monoclonal IgG, 1:600 dilution), POSTN (Abcam, ab215199, Rabbit monoclonal IgG, 1:1000 dilution), ANXA1 (Abcam, ab214486, Rabbit monocle IgG, 1:6000 dilution), HSP70 (Abcam, ab181606, Rabbit monoclonal IgG, 1:1000 dilution), HSP40 (Cell Signaling Technology, 4871T, Rabbit monoclonal IgG, 1:50 dilution), NR2F1 (Abcam, ab181137, Rabbit monoclonal IgG, 1:100 dilution) and ZEB2 (Santa Cruz Biotechnology, sc‐271984, mouse monoclonal IgG, 1:50 dilution) was performed on 4‐μm‐thick paraffin‐embedded formalin‐fixed tissue.

Techniques: Expressing, Immunohistochemistry, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Biomarker Discovery