Journal: Function

Article Title: Satellite Cell Depletion Disrupts Transcriptional Coordination and Muscle Adaptation to Exercise

doi: 10.1093/function/zqaa033

Figure Lengend Snippet: PoWeR led to higher satellite cell content in the soleus and plantaris in SC+ mice. ( A ) Representative images of satellite cell IHC from the soleus across all groups showing laminin (green), nuclei (blue), and Pax7 (red; white arrows). ( B ) Satellite cell density in the soleus. ( C ) Satellite cell density in the plantaris. Scale bar = 50 µm. Data are represented as mean ± SEM. Statistical analysis: two-way ANOVA with Tukey’s multiple comparisons test. There was a main effect for PoWeR ( P < 0.0001) and satellite cell depletion ( P < 0.0001) in both the soleus and plantaris. There was a significant interaction between PoWeR and satellite cell content in the soleus ( P < 0.001) and plantaris ( P < 0.0001). * P < 0.05, ** P < 0.01.

Article Snippet: Samples were then incubated with antimouse biotin-conjugated secondary antibody against the Pax7 primary antibody (1:1000, 115-065-205, Jackson ImmunoResearch, West Grove, PA, USA) and antirabbit secondary for laminin (1:250, A11034, Alexa Fluor 488, Invitrogen, Carlsbad, CA, USA).

Techniques:

Journal: Cell Reports Methods

Article Title: African pygmy mouse iPSCs as a model for in vitro embryogenesis, interspecies chimerism, and blastocyst complementation

doi: 10.1016/j.crmeth.2025.101293

Figure Lengend Snippet: Ablation of mouse PAX7 + cells promotes APM satellite cell formation in mouse-APM chimeras (A) Schematic of experimental design to preferentially obtain APM satellite cells in mouse-APM chimeras through elimination of host mouse PAX7 + cells during postnatal growth. (B) Representative photos of mouse-APM chimeras generated from Pax7 Cre/ERT2 ; Rosa26 LSL-DTA blastocysts. Scale bars, 1 cm. (C) Representative bright-field and fluorescence overlay images of tibialis anterior (TA) muscles from the indicated animals. Scale bars, 5 mm. (D) Immunostaining images of TA muscle cross-sections for the indicated markers in the specified chimeras. Note the presence of PAX7 + /mCHERRY + cells with and without tamoxifen treatment. Scale bars, 100 μm. (E) Representative FACS plots displaying the percentage of mCHERRY + satellite cells within the ITGA7 + satellite cell population of a tamoxifen-treated vs. a B6 chimera. (F) Graph showing quantification of the representative FACS plots shown in (E) for a larger group of chimeras. Data are shown as mean ± SD. N = 3 tamoxifen-treated chimeras and N = 8 B6 chimeras. ∗∗ p ≤ 0.01. (G) Uniform manifold approximation and projection (UMAP) based on scRNA-seq of all cell populations in tamoxifen-treated mouse-APM chimera skeletal muscles, colored by the indicated cell type. SCs, satellite cells; FAPs, fibro-adipogenic progenitors. (H) Dot plot showing individual gene expression in mouse-APM cell populations used to annotate the UMAP shown in (G). (I) UMAP of all cells from skeletal muscles of a mouse-APM chimera, colored by detection of the H2B-mCherry transcript. (J) Representative bright-field and fluorescence overlay images of FACS-purified ITGA7 + /mCHERRY + and ITGA7 + /mCHERRY − myoblasts. Scale bars, 100 μm. (K) Representative immunostaining images for the skeletal muscle differentiation marker ACTN2 in ITGA7 + /mCHERRY + myoblast-derived myotubes. Scale bars, 100 μm.

Article Snippet: Mouse: B6.Cg- Pax7 tm1(cre/ERT2)Gaka /J , The Jackson Laboratory , 017763.

Techniques: Generated, Fluorescence, Muscles, Immunostaining, Gene Expression, Purification, Marker, Derivative Assay

Journal: Experimental & Molecular Medicine

Article Title: The asymmetrical ROS–METTL3–ESR1 axis in paraspinal muscle progenitor cells determines the progression of adolescent idiopathic scoliosis

doi: 10.1038/s12276-026-01658-7

Figure Lengend Snippet: a Relative mRNA expression of METTL3 in human primary muscle stem/progenitor cells treated with or without H 2 O 2 . b , c Representative immunofluorescent staining ( b ) and relative statistical analysis ( c ) of human primary muscle stem/progenitor cells treated with or without H 2 O 2 . Green indicates METTL3; blue indicates DAPI staining of nuclei. The merged images are shown. Scale bars, 100 μm. *** P < 0.001. d The protein level of METTL3 and ESR1 in human primary muscle stem/progenitor cells treated with or without H 2 O 2 . GAPDH served as the internal reference. e Diagram illustrating the generation of Pax7-CreERT2 ; Mettl3 flox/flox mice. f , g The relative mRNA ( f ) and protein ( g ) expression level of Mettl3 in muscle stem cell isolated from Mettl3 flox/flox mice (WT) and Pax7-CreERT2 ; Mettl3 flox/flox mice (KO). h Representative immunofluorescent staining of paraspinal muscle cryosections from Mettl3 WT and KO mice. Green indicates laminin. Scale bars, 50 μm. i Statistical analysis of cross-sectional area of paraspinal muscle derived from Mettl3 WT and KO mice, respectively. At least 500 fibers were analyzed for each sample. n = 5. ** P < 0.01. j Representative immunofluorescent staining of myotubes differentiated from muscle stem cells isolated from Mettl3 WT and KO mice. Red indicates MyHC; blue indicates DAPI staining of nuclei. The merged images are shown. Scale bars, 100 μm. k Quantification of the percentage of nuclei in MyHC + cells. *** P < 0.001. l Relative mRNA expression levels of myogenic differentiation markers. Total RNA was extracted from myotubes differentiated from muscle stem cells isolated from WT and KO mice, and then RT–qPCR analysis was performed. n = 3. *** P < 0.001.

Article Snippet: The Pax7-CreERT2 mice were sourced from The Jackson Laboratory (cat. no. 011763), while the Mettl3 flox/flox mice were procured from Gem Pharmatech (cat. no. T006659).

Techniques: Expressing, Staining, Isolation, Derivative Assay, Cell Characterization, Quantitative RT-PCR

Journal: Experimental & Molecular Medicine

Article Title: The asymmetrical ROS–METTL3–ESR1 axis in paraspinal muscle progenitor cells determines the progression of adolescent idiopathic scoliosis

doi: 10.1038/s12276-026-01658-7

Figure Lengend Snippet: a Relative mRNA expression of Esr1 in muscle stem cells isolated from Pax7-CreERT2 ; Mettl3 flox/flox (WT) and Pax7-CreERT2 ; Mettl3 KO (KO) mice. ** P < 0.01. b The protein level of Esr1 and Mettl3 in muscle stem cells isolated from WT and KO mice. c Potential m 6 A modification sites on Esr1 mRNA predicted by SRAMP. d MeRIP-seq showing m 6 A peak results of Esr1 in Mettl3 flox/flox and Mettl3 -KO MuSCs. e Detailed gene sequence about the m 6 A peak of Esr1 located at chromosome 10: 5001574–5001961. The predicted functional m 6 a modification site 2,409-bp A (highlighted) was also located in this gene sequence. f MeRIP–qPCR experiment of five segments on Esr1 . *** P < 0.001. g Schematic representation of luciferase reporter assays. WT Esr1 -3′UTR, or mutant of the second potential site (Mut: AGACT to AGGCT) Esr1 -3′UTR was individually inserted behind the F-luc coding region in the luciferase reporter. h Rrelative luciferase activity. ** P < 0.01; ns indicates no significant changes. i Results of mRNA stability assay. Esr1 mRNA level were determined by RT–qPCR in muscle stem cells from WT and KO mice after actinomycin D treatment (normalized to 0 h). *** P < 0.001. j – l Results of protein stability assay. Esr1 protein levels were determined by western blot tests in muscle stem cells from WT ( j ) and KO ( k ) mice after cycloheximide treatment (normalized to 0 h) ( l ).

Article Snippet: The Pax7-CreERT2 mice were sourced from The Jackson Laboratory (cat. no. 011763), while the Mettl3 flox/flox mice were procured from Gem Pharmatech (cat. no. T006659).

Techniques: Expressing, Isolation, Modification, Sequencing, Functional Assay, Luciferase, Mutagenesis, Activity Assay, Stability Assay, Quantitative RT-PCR, Western Blot

Journal: European journal of applied physiology

Article Title: Acute resistance exercise increases the expression of chemotactic factors within skeletal muscle.

doi: 10.1007/s00421-014-2936-4

Figure Lengend Snippet: Fig. 6 Representative serial sections indicating co-locali- zation of MCP-1 with satellite cells (PAX7) and showing MCP-1 in close proximity to blood vessels (Collagen IV) 2 h following resistance exercise. Sections were stained with either anti-mouse MCP-1 (a), PAX7 (b) or Collagen IV (c), all of which appear green. Sections were also double stained with an antibody against laminin (red sarcolemma) and bisbenzamide (blue nuclei). Scale bar 100 µm (color figure online)

Article Snippet: All data were Antigen Antibody clone Dilution Source McP-1 Mouse monoclonal Igg 5J 1:20 Santa cruz (Santa cruz, cA) Il-8 Mouse monoclonal Igg nYr-HIl8 1:40 Santa cruz cD68 Mouse monoclonal Igg E-11 1:100 Santa cruz PAX7 Mouse monoclonal Igg PAX7 1:20 Developmental Studies Hybridoma Bank (Iowa city, IA) collagen IV Mouse monoclonal Igg M3F7 1:100 Developmental Studies Hybridoma Bank 1 3 checked prior to analysis to confirm that they were normally distributed.

Techniques: Staining

Journal: Developmental biology

Article Title: CRISPR-Cas13d as a molecular tool to achieve targeted gene expression knockdown in chick embryos.

doi: 10.1016/j.ydbio.2024.11.013

Figure Lengend Snippet: Figure 3. One-color CRISPR-Cas13d-mediated PAX7 knockdown. (A) A representative 498

Article Snippet: 565 Jo urn al Pr e-p roo f KEY RESOURCES TABLE Reagent or resource Source Identifier Antibodies Mouse IgG1 anti-PAX7 Developmental Studies Hybridoma Bank Cat#: pax7 RRID: AB_528428 Goat IgG anti-GFP Rockland Cat#: 600-101-215 RRID: AB_218182 Rabbit igG anti-SOX10 Millipore/Sigma Cat#: HPA068898 RRID: AB_2686054 Donkey anti-Goat IgG Alexa Fluor 647 Invitrogen Cat#: A21447 Goat anti-Mouse IgG1 Alexa Fluor 647 Invitrogen Cat#: A21240 Goat anti-Rabbit IgG Alexa Fluor 568 Invitrogen Cat#: A11011 Chemicals, Peptides, and Recombinant Proteins DAPI Thermo Fisher Cat#: D1306 Fluoromount-G SouthernBiotech Cat#: 0100-01 Experimental Models: Organisms/Strains Gallus gallus Petaluma Farms (Petaluma, CA) Fertile Rhode Island Red eggs Oligonucleotides Morpholino: Control 5’-CCTCTTACCTCAGTTACAATTTATA GeneTools Morpholino: Pax7 5’-TCCGTGCGGAGCGGGTCACCCCC GeneTools; Basch et al., 2006 Primer: gRNA3 Forward: 5’-GTAATAGCGAAGAGGCCCGC Reverse: 5’-ATAGCGGCCGCTCATCTAGA This paper; IDT Custom DNA Oligos Recombinant DNA Plasmid: pCI-H2B-RFP Betancur et al., 2010 Plasmid: pCIG Megason and McMahon, 2002 Plasmid: pCAG-memRFP This paper; Addgene Addgene # 224564 Plasmid: pCAG-Cas13d-T2A-Citrine This paper; Addgene Addgene # 224565 Plasmid: pCAG-Cas13d-T2A-GFP(1-10) This paper; Addgene Addgene # 224566 Plasmid: pCAG-memRFP-3xControlgRNA This paper; Addgene Addgene # 224567 Plasmid: pCAG-nucGFP11-3xControlgRNA This paper; Addgene Addgene # 224568 Plasmid: pCAG-3xGFP11-mem-3xControlgRNA This paper; Addgene Addgene # 224583 Plasmid: pCAG-memRFP-3xPax7gRNA This paper; Addgene Addgene # 224569 Plasmid: pCAG-nucGFP11-3xPax7gRNA This paper; Addgene Addgene # 224570 Plasmid: pCAG-3xGFP11-mem-3xPax7gRNA This paper; Addgene Addgene # 224571 Software and Algorithms Fiji Schindelin et al., 2012 RRID:SCR_002285 Zen 2 Blue Zeiss Photoshop CC Adobe Prism 10 GraphPad Jo urn al Pr e-p roo f HIGHLIGHTS • A CRISPR-Cas13d system is functional in chick embryos using a two-plasmid approach • CRISPR-Cas13d-mediated knockdown of PAX7 leads to decreased expression and function • CRISPR-Cas13d and morpholino yield comparable effects for PAX7 knockdown Jo urn al Pr e-p roo f

Techniques: CRISPR, Knockdown

Journal: Developmental biology

Article Title: CRISPR-Cas13d as a molecular tool to achieve targeted gene expression knockdown in chick embryos.

doi: 10.1016/j.ydbio.2024.11.013

Figure Lengend Snippet: Figure 4. A comparison of morpholino- and CRISPR-Cas13d-mediated PAX7 knockdown 512

Article Snippet: 565 Jo urn al Pr e-p roo f KEY RESOURCES TABLE Reagent or resource Source Identifier Antibodies Mouse IgG1 anti-PAX7 Developmental Studies Hybridoma Bank Cat#: pax7 RRID: AB_528428 Goat IgG anti-GFP Rockland Cat#: 600-101-215 RRID: AB_218182 Rabbit igG anti-SOX10 Millipore/Sigma Cat#: HPA068898 RRID: AB_2686054 Donkey anti-Goat IgG Alexa Fluor 647 Invitrogen Cat#: A21447 Goat anti-Mouse IgG1 Alexa Fluor 647 Invitrogen Cat#: A21240 Goat anti-Rabbit IgG Alexa Fluor 568 Invitrogen Cat#: A11011 Chemicals, Peptides, and Recombinant Proteins DAPI Thermo Fisher Cat#: D1306 Fluoromount-G SouthernBiotech Cat#: 0100-01 Experimental Models: Organisms/Strains Gallus gallus Petaluma Farms (Petaluma, CA) Fertile Rhode Island Red eggs Oligonucleotides Morpholino: Control 5’-CCTCTTACCTCAGTTACAATTTATA GeneTools Morpholino: Pax7 5’-TCCGTGCGGAGCGGGTCACCCCC GeneTools; Basch et al., 2006 Primer: gRNA3 Forward: 5’-GTAATAGCGAAGAGGCCCGC Reverse: 5’-ATAGCGGCCGCTCATCTAGA This paper; IDT Custom DNA Oligos Recombinant DNA Plasmid: pCI-H2B-RFP Betancur et al., 2010 Plasmid: pCIG Megason and McMahon, 2002 Plasmid: pCAG-memRFP This paper; Addgene Addgene # 224564 Plasmid: pCAG-Cas13d-T2A-Citrine This paper; Addgene Addgene # 224565 Plasmid: pCAG-Cas13d-T2A-GFP(1-10) This paper; Addgene Addgene # 224566 Plasmid: pCAG-memRFP-3xControlgRNA This paper; Addgene Addgene # 224567 Plasmid: pCAG-nucGFP11-3xControlgRNA This paper; Addgene Addgene # 224568 Plasmid: pCAG-3xGFP11-mem-3xControlgRNA This paper; Addgene Addgene # 224583 Plasmid: pCAG-memRFP-3xPax7gRNA This paper; Addgene Addgene # 224569 Plasmid: pCAG-nucGFP11-3xPax7gRNA This paper; Addgene Addgene # 224570 Plasmid: pCAG-3xGFP11-mem-3xPax7gRNA This paper; Addgene Addgene # 224571 Software and Algorithms Fiji Schindelin et al., 2012 RRID:SCR_002285 Zen 2 Blue Zeiss Photoshop CC Adobe Prism 10 GraphPad Jo urn al Pr e-p roo f HIGHLIGHTS • A CRISPR-Cas13d system is functional in chick embryos using a two-plasmid approach • CRISPR-Cas13d-mediated knockdown of PAX7 leads to decreased expression and function • CRISPR-Cas13d and morpholino yield comparable effects for PAX7 knockdown Jo urn al Pr e-p roo f

Techniques: Comparison, CRISPR, Knockdown