Journal: ecancermedicalscience

Article Title: High-risk gastrointestinal stromal tumour (GIST) and synovial sarcoma display similar angiogenic profiles: a nude mice xenograft study

doi: 10.3332/ecancer.2017.726

Figure Lengend Snippet: Supplementary. Genes and assays included in the low-density array for the study of the expression of angiogenic factors by quantitative RT-PCR.

Article Snippet: THBS4-Hs00170261_m1 , 0.25605658 , 0.041287363 , 1.055909 , 2.2569547 , 0.012994295 , , 1.6387945.

Techniques: Expressing, Amplification

Journal: ecancermedicalscience

Article Title: High-risk gastrointestinal stromal tumour (GIST) and synovial sarcoma display similar angiogenic profiles: a nude mice xenograft study

doi: 10.3332/ecancer.2017.726

Figure Lengend Snippet: Supplementary. 2- DDCt values corresponding to the Nu335 series.

Article Snippet: THBS4-Hs00170261_m1 , 0.25605658 , 0.041287363 , 1.055909 , 2.2569547 , 0.012994295 , , 1.6387945.

Techniques:

Journal: ecancermedicalscience

Article Title: High-risk gastrointestinal stromal tumour (GIST) and synovial sarcoma display similar angiogenic profiles: a nude mice xenograft study

doi: 10.3332/ecancer.2017.726

Figure Lengend Snippet: Supplementary. 2- DDCt values corresponding to the Nu407 series.

Article Snippet: THBS4-Hs00170261_m1 , 0.25605658 , 0.041287363 , 1.055909 , 2.2569547 , 0.012994295 , , 1.6387945.

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 expression in human skin following burn injury. (A) Normal skin from healthy controls; (B) skin from burn injury patients. Biopsy samples were collected from the study area at 3, 14, and 21 days after the wound excision. E—epidermis. 3 representative samples in each group are shown. Scale bar is 200 μm. (C) Relative quantification of THBS4 expression by mean integrated density of the fluorescence signal. The plot depicts the distribution of 10 samples, * indicates a statistically significant ( P < 0.05) difference.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Expressing, Fluorescence

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 is upregulated in regenerating mouse skin. Full-thickness dermal wounds were generated in mouse dorsal skin. (A) THBS4 expression was characterized by immunofluorescence microscopy in healthy skin and at 2-, 4-, 6-, and 8-days post wounding. 3 representative samples in each group are shown. Yellow arrows indicate THBS4 expression in hair follicles. Scale bar is 200 μm. (B) Relative quantification of THBS4 expression by mean integrated density of the fluorescence signal. Bars show the average of 3 samples for each time point ± standard deviation, * indicates a statistically significant ( P < 0.05) difference.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Generated, Expressing, Immunofluorescence, Microscopy, Fluorescence, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 expression in healthy control skin and psoriatic skin lesions. E—epidermis; D—dermis. 3 representative samples in each group are shown (A) and relative quantification of THBS4 expression by mean integrated density of the fluorescence signal (B) , n = 5. Scale bar is 200 μm. (C) THBS4 co-localization with fibroblast marker vimentin (Vim) and integrin beta 4 (ITGB4). Scale bar is 50 μm. The plot depicts the distribution of 5 samples, * indicates a statistically significant ( P < 0.05) difference.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Expressing, Fluorescence, Marker

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 promotes fibroblast migration. (A) Transwell migration assay with fibroblasts stimulated with recombinant THBS4 and quantification of the number of migrating cells in a field of view. (B) Transwell invasion assay through Matrigel-coated chambers and quantification of the number of migrating cells in a field of view. (C) Representative images of the in vitro scratch wound healing assay with fibroblasts stimulated with recombinant THBS4 and quantification of the relative wound closure in time. Scale bar is 200 μm. The graphs depict the averages of at least 3 independent replicates ± standard deviation, * indicates a statistically significant ( P < 0.05) difference compared to cells stimulated with control medium.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Migration, Transwell Migration Assay, Recombinant, Transwell Invasion Assay, In Vitro, Wound Healing Assay, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 promotes keratinocyte but not fibroblast proliferation in vitro . (A) Representative images of primary human fibroblasts cultured in the presence of recombinant THBS4 protein and the quantification of Ki67 + positive cells, n = 3 (B) . (C) Representative images of primary human keratinocytes cultured in the presence of recombinant THBS4 protein and quantification of Ki67 + positive cells. (D) Scale bar is 200 μm. The graphs depict the averages of at least 3 independent replicates ± standard deviation, * indicates a statistically significant ( P < 0.05) difference compared to cells stimulated with control medium.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: In Vitro, Cell Culture, Recombinant, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: Heatmap of differentially enriched genes in fibroblasts in response to THBS4 stimulation. Statistically significant ( P < 0.05) and expression fold-change > 2 protein coding genes and microRNAs are shown, n = 2.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: Ingenuity Pathway Analysis of the changes in fibroblast transcription profile in response to THBS4 stimulation. Causal network showing Frizzled and β-catenin 1 pathway activation is shown. Upper values indicate log2 fold changes, lower values show the P -value for the RNAseq.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Activation Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: Proteotranscriptomic analysis of THBS4 stimulation in fibroblasts. The combined network of transcriptomics analysis at 4 h stimulation and proteomics analysis at 24 h is shown. Single numbers indicate log2FC in proteomics and duplicate values indicate log2FC and P -value for RNAseq.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques:

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 promotes wound healing in vivo . Full dermal wounds were inflicted on the dorsal skin of C57/Bl6 mice and either recombinant THBS4 or PBS was applied. Representative images of the wounds from indicated time points (A) and measurements of the wound area (B) . The graph depicts the averages of 10 biological replicates ± standard deviation, * indicates a statistically significant ( P < 0.05) difference.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: In Vivo, Recombinant, Standard Deviation

Journal: Frontiers in Cell and Developmental Biology

Article Title: Thrombospondin-4 Is a Soluble Dermal Inflammatory Signal That Selectively Promotes Fibroblast Migration and Keratinocyte Proliferation for Skin Regeneration and Wound Healing

doi: 10.3389/fcell.2021.745637

Figure Lengend Snippet: THBS4 is a soluble dermal inflammatory signal that activates the fibroblast migration for skin regeneration and wound healing. See section “Discussion” for closer explanation.

Article Snippet: To generate THBS4 -expression plasmid, human THBS4 sequence was amplified from ORF cDNA clone expression plasmid (catalog no HGI8843-UT, Sino Biological, Beijing, China).

Techniques: Migration

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