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: European journal of immunology

Article Title: Immunomodulatory effects associated with a live vaccine against Leishmania major containing CpG oligodeoxynucleotides.

doi: 10.1002/eji.200636472

Figure Lengend Snippet: Figure 5. (A) Intracellular staining of CD3–CD11c+ cells (labeled in the figure as CD11c+) for IL-6, IL-10 and IL-12 at wk 2 post vaccination. Mice were vaccinated in the ear dermis with 104 Lm alone or in combination with 50 lg CpG, and treated with anti-IL-6R antibody (5 lg) on days –1, 1 and 3. Data are expressed as mean percentage SEM from the data obtained in three independent experiments. * Statistically significant, p = 0.008; n.s., not significant. (B) Total number of CD25hiIL-10+

Article Snippet: In vivo anti-IL-6R antibody treatment B/6mice were injected with 5 lg anti-IL-6R antibody (Cat. No. AF1830, R&D Systems, Minneapolis, MN) by intraperitoneal injection on days –1, 1 and 3 relative to vaccination with 104 Lm alone or in combination with 50 lg CpG.

Techniques: Staining, Labeling

Journal: Nature Aging

Article Title: Epigenetic dysregulation in aged muscle stem cells drives mesenchymal progenitor expansion via IL-6 and Spp1 signaling

doi: 10.1038/s43587-025-01002-0

Figure Lengend Snippet: a , Representative images of co-cultures of control or i Ezh2 −/− MuSCs and FAPs. The number of Pax7 + cells is reduced and FAPs are increased in i Ezh2 −/− MuSCs and FAPs co-cultures. The experiment was independently repeated three times with similar results. b , Bulk-RNA-seq tracks of IL-6 and Spp1 in FAPs isolated from either control or i Ezh2 −/− mice. c , EdU incorporation in co-cultures of MuSCs and PDGFRα EGFP + FAPs untreated or treated with either IL-6R or Spp1 neutralizing antibodies. MuSCs were isolated from either control PDGFRα EGFP or i Ezh2 −/− ; PDGFRα EGFP mice; FAPs were isolated from control PDGFRα EGFP mice. Arrows indicate GFP + EdU + cells. Right panel shows quantification of PDGFRα EGFP + EdU + cells. n= number of regions counted (n = 10), with cells pooled from three independent experiments. ** P = 0.0069, *** P = 0.0004 (untreated Ctrl MuSCs vs. untreated i Ezh2 −/− MuSCs), *** P = 0.0005 (untreated i Ezh2 −/− MuSCs vs. IL6R-treated i Ezh2 −/− MuSCs), ns= non-significant. d , αSMA immunostaining of co-cultured MuSCs and PDGFRα EGFP + FAPs untreated or treated with either IL-6R antibody or Spp1 neutralizing antibodies. MuSCs were isolated as described above. Right panel shows quantification of PDGFRα EGFP + αSMA + cells. n= number of regions counted (n = 10), with cells pooled from three independent experiments. * P = 0.0223 (untreated Ctrl MuSCs vs. untreated i Ezh2 −/− MuSCs), * P = 0.0169 (untreated i Ezh2 −/− MuSCs vs. IL6R-treated i Ezh2 −/− MuSCs), ** P = 0.0038, ns= non-significant. Two-way ANOVA with Sidak’s multiple comparisons ( c, d ).

Article Snippet: A solution of the anti-IL-6R monoclonal antibody (1 mg ml −1 ) (Bio X Cell, BE0047) and the mouse Spp1 polyclonal antibody (200 μg ml −1 ) (R&D Systems, AF808) were prepared in PBS shortly before the injections.

Techniques: Control, RNA Sequencing, Isolation, Immunostaining, Cell Culture

Journal: Nature Aging

Article Title: Epigenetic dysregulation in aged muscle stem cells drives mesenchymal progenitor expansion via IL-6 and Spp1 signaling

doi: 10.1038/s43587-025-01002-0

Figure Lengend Snippet: a , Representative images of Pax7 immunostaining and GFP reactivity of co-cultures with MuSCs isolated from either adult (top panel) or aged (bottom panel) (24-month-old) mice at 3 dpi and FAPs isolated from Pax7 creER ; PDGFRα EGFP adult mice at 3 dpi ( n = 3 independent experiments). b , EdU incorporation, GFP reactivity and αSMA immunostaining of co-cultured MuSCs and GFP + FAPs. Arrows indicate PDGFRα EGFP + EdU + cells. Right panel shows quantification of PDGFRα EGFP + , PDGFRα EGFP + EdU + and PDGFRα EGFP + αSMA + cells. n = number of regions counted, with cells pooled from three independent experiments ( n = 45 for PDGFRα EGFP + and PDGFRα EGFP + EdU + and n = 17 for PDGFRα EGFP + αSMA + ), **** P < 0.0001, two-tailed unpaired t -test. c , Bar plots showing IL-6 and Spp1 concentrations in the supernatants from adult, i Ezh2 −/− or aged MuSCs. Data are presented as mean ± s.d. ( n = 4 independent experiments) * P = 0.0461 (IL-6 panel) * P = 0.0321 (Spp1 panel), ** P = 0.0069 (IL-6 panel), *** P = 0.0001 (Spp1 panel). One-way ANOVA with Dunnett’s multiple comparison. d , Spp1 immunostaining of TA muscle sections from adult, i Ezh2 −/− and aged mice at 7 dpi. DAPI marks nuclei. Three muscle sections per mouse were analyzed ( n = 3 mice). e , EdU incorporation in co-cultures of MuSCs and PDGFRα EGFP + FAPs untreated or treated with either IL-6R or Spp1 neutralizing antibodies. MuSCs were isolated from either adult or aged mice; FAPs were isolated from Pax7 creER ; PDGFRα EGFP mice. Arrows indicate GFP + EdU + cells. Bottom panel shows quantification of PDGFRα EGFP + EdU + cells. n = number of regions counted, with cells pooled from three independent experiments ( n = 22) **** P = < 0.0001. f , αSMA immunostaining of co-cultured MuSCs and PDGFRα EGFP + FAPs untreated or treated with either IL-6R antibody or Spp1 neutralizing antibodies. MuSCs were isolated as described above. Right panel shows quantification of PDGFRα EGFP + αSMA + cells. n = number of regions counted, with cells pooled from three independent experiments ( n = 22) **** P = < 0.0001, NS, non-significant. Two-way ANOVA with Sidak’s multiple comparisons ( e , f ). αIL6R, against-IL6R antibody; αSppl1, against-Spp1 antibody.

Article Snippet: A solution of the anti-IL-6R monoclonal antibody (1 mg ml −1 ) (Bio X Cell, BE0047) and the mouse Spp1 polyclonal antibody (200 μg ml −1 ) (R&D Systems, AF808) were prepared in PBS shortly before the injections.

Techniques: Immunostaining, Isolation, Cell Culture, Two Tailed Test, Comparison

Journal: Nature Aging

Article Title: Epigenetic dysregulation in aged muscle stem cells drives mesenchymal progenitor expansion via IL-6 and Spp1 signaling

doi: 10.1038/s43587-025-01002-0

Figure Lengend Snippet: a , Schematic of IL-6R and Spp1 antibodies injection in aged (24-month-old) mice. b , Pdgfrα immunostaining of TA muscle sections from adult or aged mice injected with either PBS or Spp1 and IL-6R antibodies at 14 dpi. DAPI identifies cell nuclei. On the right is the quantification of Pdgfrα + cells. Two muscle sections per mouse were analyzed. Data are presented as mean ± s.d., two-tailed unpaired t -test, n = 4 mice, * P = 0.0425. c , Representative images of Spp1 immunostaining of TA muscle sections from aged mice injected with either PBS or Spp1 and IL-6R antibodies at 14 dpi. DAPI identifies cell nuclei. The experiment was independently repeated three times with similar results. d , Laminin immunostaining of TA muscle sections from aged mice treated with PBS (untreated) or with Spp1 and IL-6R antibodies at 14 dpi. DAPI identifies cell nuclei. Right panel shows quantification of myofiber CSA analyzed. Two muscle sections per mouse were analyzed. Data are presented as mean ± s.d., two-tailed unpaired t -test. n = 8 TAs from four PBS-treated or against-Spp1 antibody-treated and against-IL6R antibody-treated mice, respectively, * P = 0.0197 (0−200-μm 2 fibers). e , Picrosirius Red staining and quantification of fibrotic area in TA muscle sections from aged mice injected with PBS or with Spp1 and IL-6R antibodies at 14 dpi. Two muscle sections per mouse were analyzed. Data are presented as mean ± s.d., two-tailed unpaired t -test. n = 6 TA from three PBS-treated or against-Spp1 antibody-treated and against-IL-6R antibody-treated mice-treated mice, respectively, * P = 0.0338. αIL6R, against-IL6R antibody; αSppl1, against-Spp1 antibody.

Article Snippet: A solution of the anti-IL-6R monoclonal antibody (1 mg ml −1 ) (Bio X Cell, BE0047) and the mouse Spp1 polyclonal antibody (200 μg ml −1 ) (R&D Systems, AF808) were prepared in PBS shortly before the injections.

Techniques: Injection, Immunostaining, Two Tailed Test, Staining