Journal: PloS one

Article Title: Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans.

doi: 10.1371/journal.pone.0006027

Figure Lengend Snippet: Figure 1. Circulating IL-6 and muscle IL-6 mRNA and IL-6 receptor response muscle lengthening contractions (MLC). (1a) Average serum interleukin-6 (IL-6) concentration. Time 0 hr corresponds to pre-intervention values; all other time-points correspond to post-intervention time (hr). (1b) Relative IL-6 mRNA expression, expressed as fold-change from 0 hr. (1c) Pearson correlation of serum IL-6 concentration versus muscle IL-6 mRNA (fold-change), correlation is representative of the individual data points and presented as mean values ($)6SD (error bars). (1d) Relative IL-6 receptor (IL-6Ra) mRNA expression, expressed as fold-change from 0 hr. (1e) Immunofluorescent image (406) of a muscle cross-section triple-stained for Pax7 (red), IL-6Ra (green) and nuclei (DAPI = blue); IL-6Ra staining is apparent on the sarcolemma and satellite cell membrane. White arrow denotes one of the Pax7+ nuclei which co-localized with IL-6Ra (scale bar = 100 mm). Values are reported as mean6S.E.M. Mean values represent the mean for all 8 subjects per time-point (8 samples per time-point, 40 samples total). *p,0.05 vs. 0 hr. doi:10.1371/journal.pone.0006027.g001

Article Snippet: Immunofluorescence 7 mm sections were cryosectioned and stained with antibodies against Pax7 (neat; DSHB, USA); IL-6 (500 ng/mL, MAB 2061, R&D Systems, USA); p-STAT3 (p-STAT3 Y705 1:100, Cell Signaling Technologies Inc., USA); IL-6Ra (1:50, MCA822, Serotec, UK); PCNA (1:200, ab15497, Abcam Inc., USA); and Laminin (1:1000, L8271, Sigma-Aldrich, Canada).

Techniques: Concentration Assay, Expressing, Staining, Membrane

Journal: Scientific Reports

Article Title: Recombinant soluble gp130 protein reduces DEN-induced primary hepatocellular carcinoma in mice

doi: 10.1038/srep24397

Figure Lengend Snippet: ( a ) Schematic diagrams of sgp130 in inhibiting IL-6 signaling. ( b ) Expression of p-STAT3, IL-6, IL-6R and gp130 in mouse liver tissue (400×). (A: Control; B: model group; C: sgp130 treatment group). ( c ) sgp130 changes the DEN-induced high level of liver related factors in mouse sera. ( d ) B-ultrasonography of liver for detecting DEN-induced hepatic fibrosis (Arrowheads point to hepatic fibrosis bright spots or stripes; (A: Control; B: model group; C: sgp130 treatment group)). ( e ) type IV collagen expressed in tissue of mouse livers, detected with immunohistochemical staining (400×) (A: Control; B: model group; C: sgp130 treatment group).

Article Snippet: Primary specific Anti-p-STAT3 antibody was purchased from Santa Cruz, CA, USA and specific antibodies to Ki67, PCNA, PSA, IL-6, IL-6R, gp130, EGFR and TGFβ were gifts from Maixin biotech., Fuzhou, China.

Techniques: Expressing, Control, Immunohistochemical staining, Staining

Journal: Scientific Reports

Article Title: Recombinant soluble gp130 protein reduces DEN-induced primary hepatocellular carcinoma in mice

doi: 10.1038/srep24397

Figure Lengend Snippet: ( a ) The xenograft tumors and their curve on nude mice receiving a subcutaneous injection of 5 × 10 6 /mL HepG2 cells. Data are mean ± SD, n = 6 (Single*, double** and triple*** marked represent statistical significance in p < 0.05, p < 0.01 and p < 0.001, respectively). Each experiment was repeated three times, and data shown are representative one of three independent experiments. ( b ) Immunohistochemical analysis of IL-6, IL-6R, gp130, Ki67 and PCNA expression in xenograft tumors (100×) (A: Control; B: sgp130 treatment group). ( c ) Detection of HepG2 cell metastasis in nude mouse lungs through analyzing expression of CD44 and Ki67 lung tissues (100X) (A: Negative control; B: Control; C: sgp130 treatment group).

Article Snippet: Primary specific Anti-p-STAT3 antibody was purchased from Santa Cruz, CA, USA and specific antibodies to Ki67, PCNA, PSA, IL-6, IL-6R, gp130, EGFR and TGFβ were gifts from Maixin biotech., Fuzhou, China.

Techniques: Injection, Immunohistochemical staining, Expressing, Control, Negative Control

Journal: Scientific Reports

Article Title: Recombinant soluble gp130 protein reduces DEN-induced primary hepatocellular carcinoma in mice

doi: 10.1038/srep24397

Figure Lengend Snippet: the sequence of primers.

Article Snippet: Primary specific Anti-p-STAT3 antibody was purchased from Santa Cruz, CA, USA and specific antibodies to Ki67, PCNA, PSA, IL-6, IL-6R, gp130, EGFR and TGFβ were gifts from Maixin biotech., Fuzhou, China.

Techniques: Sequencing

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. qPCR results from pig articular chondrocytes stimulated with OSM and treated with or without SRC inhibitor (SU6656) for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein expression of pSRC in ATDC5 cells after transfection (72 hours) with the modified and WT gp130 plasmids stimulated with OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. qPCR results from ATDC5 cells treated with SRC inhibitor (SU6656) alone, or transfected with indicated variants of plasmids and stimulated with or without OSM for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. D. Protein expression of gp130 pY814 in untreated fetal (17 weeks) and adult articular chondrocytes. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Transfection, Modification

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of pSRC in pig articular chondrocytes stimulated with IL-6 cytokines. Horizontal lines with bars show the mean ± SD. n=3. B. Schematic of modified amino acids within gp130 812-827 domain. Protein expression of pSRC in Ba/F3 cells after transfection with the modified or WT gp130 plasmids stimulated with IL-6 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. CRISPR gRNA was designed by PNA Bio. Y814 mutant mouse (F814) was generated by the USC transgenic mouse core. Mice genotyping was performed by GeneWiz. Representative images of 4-month-old males shown. n=4. D. Protein expression of gp130 pY814 in wild type (WT) and F814 mouse splenocytes treated with and without OSM and LIF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. E. Distribution of differentially expressed genes in MA plot of wild type (WT) and F814 mouse periarticular stromal cells. MA plots (log2fold change vs log2mean expression) for WT-OSM vs WT and Y814-OSM vs Y814 mouse periarticular stromal cells. Each dot represents a gene. Red dots represent upregulated genes i.e. log2fold-change >1 and p-value<0.05. Blue dots represent downregulated genes i.e. log2fold change < −1 and p-value 0.05. Genes that do not qualify this threshold are indicated by grey dots. F. Levels of protein complex formation between gp130 and pSRC in wild type and F814 mouse splenocytes stimulated with or without OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. G. Protein expression of pSRC in wild type and F814 mouse splenocytes stimulated with or without OSM for 24 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Modification, Transfection, CRISPR, Mutagenesis, Generated, Transgenic Assay

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Protein expression of gp130 pY814 in wild type (WT) and F814 mouse splenocytes treated with and without OSM and LIF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein levels of pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. C. Computational analysis using GOLD software predicted a high affinity binding of peptide QQpYF to the regulatory site of SRC (c-SRC). c-SRC as visualized by crystallography. The structure of the indicated c-SRC domains is shown in ribbon diagram representation (left) as well as with electrostatic potential (blue, positive charge; red, negative charge; white, neutral) mapped onto the molecular surface (right). Peptide QQpYF is shown in stick representation. D. Transcription of genes was determined via qPCR in human adult OA articular chondrocytes treated with or without peptide QQpYF for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. E. Pig knee cartilage explants were stimulated with or without OSM and treated with or without peptide QQpYF at indicated doses for 72 hours followed by a neoepitope assay. Levels of cleaved ACAN and COL2 neoepitopes in the supernatant were quantified with respect to the wet weight of the explant. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Software, Binding Assay

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Protein levels of pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of control scrambled peptide or peptide QQpYF for 4 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques:

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of gp130 pY814 in pig articular chondrocytes treated with or without OSM and R805 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3. B. Gross morphology and Masson’s trichrome staining of bearded dragon treated with vehicle control and R805 14 days post-amputation. Dashed lines mark amputation planes. n=10. sc=spinal cord; we=wound epithelium; ve=vertebra. C. Gross morphology of bearded dragon co-treated with vehicle control or R805 and PBS or clodronate liposomes 14 days post-amputation. Dashed lines mark amputation planes. n=10. D. Mouse vehicle-treated or R805-treated post-wound day (PWD) 21 wound sections. Representative images are shown. n=8. E. Hair follicle (n=3) and fiber length (n=8) in R805-treated mouse PWD 14 and untreated control wounds. Horizontal lines with bars show the mean ± SD. F. Re-clustering of clusters annotated as macrophages from skin wounds of vehicle-treated or R805-treated mice PWD 14. Dot plots depict gene expression in each macrophage cluster. The contribution of each sample to each cluster is shown as a stacked bar graph. Dot sizes are proportional to the percentage of cells in each cluster expressing the indicated gene. DCs = dendritic cells. G. Re-clustering of clusters annotated as fibroblasts from skin wounds of vehicle-treated or R805-treated mice PWD 14. Dot plots depict gene expression in each fibroblast cluster. The contribution of each sample to each cluster is shown as a stacked bar graph.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Receptor-competition assay. Adult human articular chondrocytes were treated with or without indicated IL-6 family of cytokines and different doses of R805 for 4 hours. Co-IP measuring protein expression was performed after transfection with gp130 FLAG plasmid (72 hours) followed by western blot with respective receptor antibodies. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Competitive Binding Assay, Co-Immunoprecipitation Assay, Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Receptor-competition assay. Adult human articular chondrocytes were treated with or without indicated IL-6 family of cytokines and different doses of R805 for 24 hours. Co-IP measuring protein expression was performed after transfection with gp130 FLAG plasmid (72 hours) followed by western blot with respective receptor antibodies. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Competitive Binding Assay, Co-Immunoprecipitation Assay, Expressing, Transfection, Plasmid Preparation, Western Blot

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: A. Protein expression of gp130 pY814 in pig articular chondrocytes treated with or without OSM and R805 for 4 hours. Horizontal lines with bars show the mean ± SD. n=3. B. qPCR results from pig articular chondrocytes cells treated with OSM with or without R805 for 48 hours. Horizontal lines with bars show the mean ± SD. n=3. C. Pig cartilage knee explants were incubated with or without OSM for 72 hours with or without R805 at indicated concentrations followed by a neoepitope assay. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: Expressing, Incubation

Journal: bioRxiv

Article Title: Signaling modality within gp130 receptor enhances tissue regeneration

doi: 10.1101/2022.01.05.475124

Figure Lengend Snippet: Levels of complex formation between gp130 and pSRC in pig articular chondrocytes stimulated with or without OSM in presence or absence of R805 for 24 hours. Horizontal lines with bars show the mean ± SD. n=3.

Article Snippet: The immune complexes were sedimented, washed and separated by SDS-PAGE (see below) and further analyzed by Western blot using p-SRC (pY416-SRC) (cat# 1246F, Novus Biologicals) or NEMO (cat # 18474-1-AP, Proteintech) antibodies and normalized to gp130 (cat #bs-1459R, Bioss) or total SRC (cat # 2123, Cell Signaling).

Techniques: