(A) Schematic of experiment, whereby young (3 mo) mice were administered 1-mm (administered to left leg) or 2-mm (administered to right leg) biopsy punches to their tibialis anterior muscle to …
(A, B) Force frequency of muscle (A) and nerve stimulation (B) of uninjured, 1-mm and 2-mm groups uninjured (blue) tibialis anterior muscle at 28 d post injury (dpi) (1 mm [black] or 2 mm [gray]). …
(A) Mice were administered a bilateral defect to tibialis anteriors (TAs) (left leg received a 1-mm defect and right leg received a 2-mm defect). Tissues were harvested at 0, 3, 7, and 14 d post …
(A) Dirichlet Process Gaussian Process (DPGP) mixture model-based clustering of mean analyte abundance fold change z-scores across injury timepoints. Black lines are cluster means, and the gray bars …
(A) Mice were administered bilateral 2-mm VML defects to their tibialis anterior (TA) muscles. One limb received an intramuscular injection of Maresin 1 and the other limb received vehicle (saline + …
(A) Complete gating strategy for neutrophils and macrophages. Values indicate percentage of parent. (B) Maresin 1 treatment significantly reduced neutrophil abundance and reduced F4/80+ macrophage …
(A) Representative cross-sections from muscles isolated 7 d post injury (dpi) with vehicle (top) or Maresin 1 (bottom) treatment. Sections are immunostained for embryonic myosin heavy chain (eMyHC) …
(A, B) Force frequency of nerve (A) and muscle stimulation (B) of uninjured (blue) tibialis anterior muscle at 28 d post injury (dpi) following treatment with saline (black) or Maresin 1 (gray). …
(A) Lgr6 is highly upregulated in proliferating myoblasts by RT-qPCR. Graph shows mean ± SEM, ****p<0.0001 by one-way ANOVA and Benjamini–Hochberg (BH) post hoc analysis. n = 3–4 wells per …
(A) Maresin 1 dose–response curve showed the most impact on proliferation with treatment using 10 nM. There was no statistically significant effect with higher doses. *p<0.05 by one-way ANOVA with …
(A) Uniform Manifold Approximation and Projection (UMAP) plots colored by clusters at a resolution of 0.2 and annotated into cell types. (B) UMAP plots split by treatment and colored by cell type. (C…
(A) Uniform Manifold Approximation and Projection (UMAP) plots colored by the number of genes per cell (left) and the number of unique molecular identifiers per cell (right). (B) Dot plot of known …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Biological sample (Mus musculus) | C57BL/6J wild-type mice | Jackson Labs | Jackson Stock 000664 | Female (2–3 mo) |
Biological sample (M. musculus) | Pax7CreERT2/+;Rosa26Tcf/Lef-LSL-H2B-GFP mice | University of Michigan | Jackson stock 017763 crossed with Jackson stock 032577 | Female (2–3 mo) |
Biological sample (M. musculus) | Pax7CreERT2 – Rosa26-TdTomato | University of Michigan | Jackson stock 017763 crossed with Jackson stock 007914 | Female (2–3 mo) |
Antibody | AF488 anti-mouse monoclonal F4/80, clone: BM8, isotype: Rat IgG2a, κ | BioLegend | 123119; RRID:AB_893491 | FC (1:200) |
Antibody | APC-Cy7 anti-mouse monoclonal Ly-6G, clone: 1A8, isotype: Rat IgG2a, κ | BioLegend | 127624; RRID:AB_10640819 | FC (1:400) |
Antibody | APC anti-mouse monoclonal Ly-6A/E (Sca-1), clone: D7, isotype: Rat IgG2a, κ | BioLegend | 108112; RRID:AB_313349 | FC (1:400) |
Antibody | APC anti-mouse monoclonal CD45, clone: 30-F11, isotype: Rat IgG2b, κ | BioLegend | 103112; RRID:AB_312977 | FC (1:400) |
Antibody | APC anti-mouse monoclonal TER-119, clone: TER-119, isotype: Rat IgG2b, κ | BioLegend | 116212; RRID:AB_313713 | FC (1:400) |
Antibody | APC anti-mouse monoclonal CD31, clone: 390, Isotype: Rat IgG2a, κ | BioLegend | 102410; RRID:AB_312905 | FC (1:400) |
Antibody | APC anti-mouse/human monoclonal CD11b, clone: M1/70. Isotype: Rat IgG2b, κ | BioLegend | 101212; RRID:AB_312795 | FC (1:400) |
Antibody | PE anti-mouse/rat monoclonal CD29 (B1 int), clone: HMβ1-1, isotype: Armenian Hamster IgG | BioLegend | 102208; RRID:AB_312885 | FC (1:200) |
Antibody | Biotin Rat AntiMouse monoclonal CD184, clone: 2B11/CXCR4 (RUO), isotype: Rat IgG2b, κ | BD Biosciences | 551968; RRID:AB_394307 | FC (1:200) |
Antibody | Anti-mouse monoclonal CD68, clone: FA-11, isotype: IgG2a | Bio-Rad | MCA1957 | IF (1:50) |
Antibody | Mouse monoclonal Anti-human MYH3 | DHSB | F1.652 | IF (1:20) |
Antibody | Chicken polyclonal Anti-mouse GFP | Abcam | ab13970; RRID:AB300798 | IF (1:1000) |
Antibody | Rabbit anti-mouse laminin 1+2, Isotype: Polyclonal IgG | Abcam | ab7463; RRID:AB_305933 | IF (1:500) |
Antibody | Rabbit polyclonal Anti-RFP | Rockland | 600-401-379; RRID:AB_2209751 | IF (1:50) |
Antibody | Rabbit polyclonal Anti-Beta catenin polyclonal (CAT-15) | Thermo Fisher | 71-2700; RRID:AB_2533982 | IF (1:100) |
Antibody | Goat polyclonal Anti-rabbit IgG (H+L), Alexa Fluor 647 conjugate | Thermo Fisher | A27040; RRID:AB_2536101 | IF (1:500) |
Antibody | Goat polyclonal Anti-rat (H+L), Alexa Fluor 647 conjugate | Thermo Fisher | A21247; RRID:AB_141778 | IF (1:500) |
Antibody | Goat polyclonal Anti-mouse (H+L), Alexa Fluor 488 conjugate | Thermo Fisher | A28175; RRID:AB_2536161 | IF (1:500) |
Antibody | Goat polyclonal Anti-chicken IgY (H+L), Alexa Fluor 488 conjugate | Thermo Fisher | A32931TR; RRID:AB_2866499 | IF (1:500) |
Antibody | Goat polyclonal Anti-rabbit (H+L), Alexa Fluor 555 conjugate | Thermo Fisher | A21247; RRID:AB_141778 | IF (1:500) |
Sequence-based reagent | PrimeTime Mouse GAPDH Primer | Integrated DNA Technologies | Mm.PT.39a1 | |
Sequence-based reagent | PrimeTime Mouse Lgr6 Primer | Integrated DNA Technologies | Mm.PT.58.9348010 | |
Sequence-based reagent | Mouse Lgr6 DsiRNA #1 | Integrated DNA Technologies | mm.Ri.Lgr6.13.2 | |
Sequence-based reagent | Mouse Lgr6 DsiRNA #2 | Integrated DNA Technologies | mm.Ri.Lgr6.13.1 | |
Sequence-based reagent | Cell Multiplexing Oligos | 10x Genomics | 1000261 | |
Peptide, recombinant protein | Bovine serum albumin | Fisher Scientific | BP9703-100 | |
Peptide, recombinant protein | Fibroblast growth factor basic | Gibco-Invitrogen | PHG0263 | |
Peptide, recombinant protein | 0.25% Trypsin EDTA | Gibco-Invitrogen | 25200072 | |
Peptide, recombinant protein | HALT Protease Inhibitor Cocktail | Thermo Fisher | 87786 | |
Chemical compound, drug | Dispase II (activity ≥ 0.5 units/mg solid) | Sigma | D4693-1G | Digestion of tissue to extract MuSCs |
Chemical compound, drug | Collagenase type II (654 U/mg, non-specific proteolytic activity 487 U/mg) | Life Technologies | 17101015 | Digestion of tissue to extract MuSCs |
Chemical compound, drug | DMEM, high glucose, pyruvate | Life Technologies | 11995065 | |
Chemical compound, drug | Ham’s F-10 Nutrient Mix | Life Technologies | 11550043 | |
Chemical compound, drug | Tissue Plus O.C.T Compound | Fisher Scientific | 23-730-571 | |
Chemical compound, drug | Magnesium sulfate heptahydrate | Sigma-Aldrich | 63138-250G | |
Chemical compound, drug | Sodium bicarbonate | Sigma-Aldrich | S5761 | |
Chemical compound, drug | SafeClear II | Fisher Scientific | 23-044192 | |
Chemical compound, drug | Picric acid | Sigma-Aldrich | P6744-1GA | |
Chemical compound, drug | Glacial acetic acid | Sigma-Aldrich | BP2401-500 | |
Chemical compound, drug | Xylenes | Sigma-Aldrich | 534056-4L | |
Chemical compound, drug | Permount | Fisher Scientific | SP15-100 | |
Chemical compound, drug | 0.5 M EDTA | Invitrogen | 15575-038 | |
Chemical compound, drug | Sodium azide | Sigma-Aldrich | 71289 | |
Chemical compound, drug | 4% paraformaldehyde in PBS | Thermo Fisher | J19943-K2 | |
Chemical compound, drug | Penicillin streptomycin | Gibco-Invitrogen | 15640055 | |
Chemical compound, drug | Maresin 1 | Cayman Chemicals | 10878 | |
Chemical compound, drug | Tween-20 | Sigma-Aldrich | P1379 | |
Chemical compound, drug | TritonX-100 | Sigma-Aldrich | T8787 | |
Chemical compound, drug | Tamoxifen | Sigma-Aldrich | T2859-1G | |
Chemical compound, drug | Corn Oil | Sigma-Aldrich | C8267-2.5L | |
Commercial assay or kit | Satellite Cell Isolation Kit, mouse | Miltenyi | 130-104-268 | |
Commercial assay or kit | TGF beta 1 DuoSet ELISA Kit | R&D Systems | DY1679 | |
Commercial assay or kit | Pierce BCA Assay Kit | Thermo Fisher | 23227 | |
Commercial assay or kit | EdU ClickIt Reaction Kit | Thermo Fisher | C10340 | |
Commercial assay or kit | SuperScript III First-Strand Synthesis Kit | Thermo Fisher | 18080051 | |
Commercial assay or kit | QIAGEN RNeasy Mini Kit | QIAGEN | 74104 | |
Commercial assay or kit | Qubit RNA HS Assay | Thermo Fisher | Q32852 | |
Commercial assay or kit | Single cell 3’ Library & Gel Bead Kit | 10x Genomics | 120267 | |
Commercial assay or kit | cAMP-Glo Assay | Promega | V1501 | |
Software, algorithm | CellRanger v7.0.0 | 10x Genomics | https://support.10xgenomics.com/single-cell-gene-expression/software/downloads | |
Software, algorithm | R v4.2.1 | The R Foundation for Statistical Computing | https://www.r-project.org/; RRID:SCR_001905 | |
Software, algorithm | Python | https://www.python.org/ | ||
Software, algorithm | MetaboDiff v0.9.5 | Mock et al., 2018; Mock, 2020 | https://github.com/andreasmock/MetaboDiff | |
Software, algorithm | DPGP v0.1 | McDowell et al., 2018a; McDowell et al., 2018b | https://github.com/PrincetonUniversity/DP_GP_cluster | |
Software, algorithm | MATLAB_R2020a | MathWorks | https://www.mathworks.com/products/matlab.html | |
Software, algorithm | Seurat v4.2.1 | Stuart et al., 2019 | https://satijalab.org/seurat/; RRID:SCR_007322 | |
Software, algorithm | ggplot2 v3.2.1 | Wickham et al., 2016 | https://ggplot2.tidyverse.org; RRID:SCR_014601 | |
Software, algorithm | FlowJo v10 | https://www.flowjo.com; RRID:SCR_008520 | ||
Software, algorithm | ImageJ v2.1.0 | https://imagej.net/ImageJ; RRID:SCR_002285 | ||
Software, algorithm | Cellpose | Stringer et al., 2021 | RRID:SCR_021716 | |
Software, algorithm | LabelsToROIs | Waisman et al., 2021 | https://labelstorois.github.io/ | |
Software, algorithm | Bioinformatics analysis code | This paper | https://github.com/AguilarLab/Maresin1 (copy archived at Castor-Macias et al., 2023) | Algorithm used to generate Figure 5, Figure 5—figure supplement 1. Refer to data processing and analysis under single-cell RNA sequencing |
Other | Streptavidin PE-Cyanine7 | Thermo Fisher | 25-4317-82; RRID:AB_10116480 | FC (1:100) |
Other | scRNA-seq datasets | This paper | GSE215808 | Refer to data processing and analysis under single-cell RNA sequencing. |
Other | LS Columns | Miltenyi | 130-042-401 | MACS isolation |
Other | UltraComp eBeads | Fisher Scientific | 01-2222-42 | Single color controls for Flow Cytometry |
Other | RIPA Buffer | Thermo Fisher | 89900 | Tissue homogenization reagent in ELISA assays |
Other | Lipofectamine RNAiMAX | Invitrogen | 13-778-030 | Transfection reagent |
Other | Hoechst 33342 | Thermo Fisher | H3570 | IF (1:1000) nuclear stain |
Other | SYBR Green PCR MasterMix | Thermo Fisher | 4309155 | PCR reagent |
Other | Matrigel | BD Biosciences | 356234 | Cell culture coating reagent |
Other | Horse Serum | Gibco-Invitrogen | 26050088 | Cell culture |
Other | Prolong Diamond | Thermo Fisher | P36965 | Mounting media |
Other | Direct Red 80 | Fisher Scientific | AAB2169306 | Picrosirius red stain |
Other | Hematoxylin | Ricca Chemical Company | 3530-16 | H&E stain |
Other | Eosin | EMD-Millipore | 588X-75 | H&E stain |
Other | Fetal bovine serum | Life Technologies | 10437028 | Tissue digestion reagent |
Other | Normal goat serum | Abcam | Ab7481; RRID:AB_2716553 | IHC stain |
Other | Propidium iodide – 1.0 mg/mL solution in Water | Life Technologies | P3566 | FC (1:400) |
Other | 7-AAD | BioLegend | 420403 | FC (1:200) |
Other | Mouse on Mouse blocking reagent | Vector Labs | MKB-2213 | IHC stain |
Metabolite concentration (pmol/g) for different pathways in uninjured, 1 mm VML, and 2 mm (TA) tissue homogenates.