myod polyclonal antibody  (Proteintech)

Journal: Frontiers in Veterinary Science

Article Title: miR-379-5p retards the proliferation and differentiation of goat skeletal muscle satellite cells by targeting LIN28B

doi: 10.3389/fvets.2025.1694160

Figure Lengend Snippet: miR-379-5p inhibits differentiation and promotes mitochondrial function in differentiating goat MuSCs. (A) Relative expression level of miR-379-5p in goat MuSCs transfected with NC or miR-379-5p mimics during the differentiation phase. (B) Relative mRNA levels of differentiation marker genes ( MyoD , MyoG , MyHC , Myf5 , and MEF2C ) in MuSCs transfected with NC or miR-379-5p mimics. (C) Western blot analysis of MyoD protein levels in differentiating MuSCs transfected with NC or miR-379-5p mimics. β-Tubulin was used as a loading control. (D) Representative immunofluorescence images of MyHC and DAPI in differentiating goat MuSCs after miR-379-5p overexpression. The right panel shows quantification of the fusion index. Scale bar = 50 μm. (E) Relative mRNA levels of mitochondrial marker genes ( CYCS , ATP5a1 , TFAM , and COX1 ) in differentiating MuSCs transfected with NC or miR-379-5p mimics. (F) Representative fluorescence images and quantification of mitochondrial membrane potential and ROS levels in differentiating MuSCs. Left: JC-1 staining reflects mitochondrial membrane potential. ROS levels were indicated by green fluorescence. Middle: Ratio of fluorescence intensity. Right: Quantification of ROS fluorescence intensity. Scale bar = 500 μm. Each experiment contained three biological replicates. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included Pax7 (1:200, Santa Cruz Biotechnology, China), PCNA (1:200, Santa Cruz Biotechnology, China), β -Tubulin mouse mAb (1:5000, ZenBioScience, China), and MyoD polyclonal antibody (1:1000, Proteintech, United States).

Techniques: Expressing, Transfection, Marker, Western Blot, Control, Immunofluorescence, Over Expression, Fluorescence, Membrane, Staining

Journal: Frontiers in Veterinary Science

Article Title: miR-379-5p retards the proliferation and differentiation of goat skeletal muscle satellite cells by targeting LIN28B

doi: 10.3389/fvets.2025.1694160

Figure Lengend Snippet: LIN28B promotes differentiation and impairs mitochondrial function in differentiating goat MuSCs. (A) Relative mRNA level of LIN28B in goat MuSCs transfected with pCtrl or pLIN28B during the differentiation phase. (B) Relative mRNA levels of differentiation marker genes ( MyoD , MyoG , MyHC , MEF2C , and Myf5 ) in MuSCs transfected with pCtrl or pLIN28B. (C) Western blot analysis of MyoD protein levels in differentiating MuSCs transfected with pCtrl or pLIN28B. β-Tubulin was used as a loading control. (D) Representative immunofluorescence images of MyHC and DAPI in differentiating MuSCs transfected with pCtrl or pLIN28B. The right panel shows quantification of the fusion index. Scale bar = 100 μm. (E) Relative mRNA levels of mitochondrial marker genes ( CYCS , ATP5a1 , TFAM , COX1, and COX2 ) in differentiating MuSCs transfected with pCtrl or pLIN28B. (F) Representative fluorescence images and quantification of mitochondrial membrane potential and ROS levels in differentiating MuSCs. Left: JC-1 staining reflects mitochondrial membrane potential. ROS levels were indicated by green fluorescence. Middle: Ratio of fluorescence intensity. Right: Quantification of ROS fluorescence intensity. Scale bar = 500 μm. Each experiment contained three biological replicates. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included Pax7 (1:200, Santa Cruz Biotechnology, China), PCNA (1:200, Santa Cruz Biotechnology, China), β -Tubulin mouse mAb (1:5000, ZenBioScience, China), and MyoD polyclonal antibody (1:1000, Proteintech, United States).

Techniques: Transfection, Marker, Western Blot, Control, Immunofluorescence, Fluorescence, Membrane, Staining

Journal: Frontiers in Veterinary Science

Article Title: miR-379-5p retards the proliferation and differentiation of goat skeletal muscle satellite cells by targeting LIN28B

doi: 10.3389/fvets.2025.1694160

Figure Lengend Snippet: miR-379-5p inhibits goat MuSC differentiation and improves mitochondrial function by targeting LIN28B . (A) Relative mRNA levels of MyoD , MyoG , and MyHC in goat MuSCs transfected with NC or miR-379-5p mimics in combination with pCtrl or pLIN28B during the differentiation phase. (B) Western blot analysis of MyoD protein levels in differentiating MuSCs transfected with NC or miR-379-5p mimics in combination with pCtrl or pLIN28B. β-Tubulin was used as a loading control. (C) Immunofluorescence staining of MyHC and analysis of myotube fusion index in differentiating goat MuSCs. Scale bar = 100 μm. (D) Relative mRNA levels of mitochondrial marker genes ( TFAM and COX1 ) in differentiating MuSCs transfected with NC or miR-379-5p mimics in combination with pCtrl or pLIN28B. (E) Representative fluorescence images illustrating mitochondrial membrane potential and ROS levels in differentiating MuSCs. Left: JC-1 staining indicates mitochondrial membrane potential. ROS production is indicated by green fluorescence. Middle: Ratio of fluorescence intensity. Right: Quantification of ROS fluorescence intensity. Scale bar = 500 μm. Each experiment contained three biological replicates. * p < 0.05, ** p < 0.01.

Article Snippet: Primary antibodies included Pax7 (1:200, Santa Cruz Biotechnology, China), PCNA (1:200, Santa Cruz Biotechnology, China), β -Tubulin mouse mAb (1:5000, ZenBioScience, China), and MyoD polyclonal antibody (1:1000, Proteintech, United States).

Techniques: Transfection, Western Blot, Control, Immunofluorescence, Staining, Marker, Fluorescence, Membrane

Journal: Cell & Bioscience

Article Title: Generation of musculoskeletal cells from human urine epithelium-derived presomitic mesoderm cells

doi: 10.1186/s13578-024-01274-w

Figure Lengend Snippet: Differentiation of UiPSM cells into skeletal myocytes in vitro. A Schematic diagram of somite development. a. Illustration of the epithelial somite's spatial relationship to surrounding structure. b. Depiction of the differentiated somite's spatial relationship to surrounding structures. Dorsally, the somite differentiates into the dermomyotome and sclerotome. The dermomyotome subsequently gives rise to the myotome, which develops into skeletal muscle tissue. The sclerotome differentiates into osteoblasts and chondroblasts, forming the axial skeleton. B Schematic diagram of UiPSM cell differentiation into skeletal myocytes, osteoblasts and chondroblasts. C Schematic overview of stepwise differentiation of skeletal myocytes from UiPSM cells. Representative images show the morphological changes from UiPSM cells to skeletal muscle filaments. Scale bars, 100 µm. D Representative gene expression of human skeletal muscle satellite cells ( PAX3, PAX7, CXCR4, C-MET ) at day 15. Data are mean ± SD, n = 3 independent experiments. (*P ≤ 0.05). E Immunofluorescence of PAX3 and PAX7 during the differentiation of skeletal muscle cell form UiPSM at day 60 (left). The scale bar represents 100 µm. The values on the left represent the percentage of positive cells statistically. Data are mean ± SD, n = 3 independent experiments, each experiment counted 100 fields of view. F Representative gene expression of human skeletal muscle satellite cells ( PAX3, PAX7 ) and skeletal myoblasts ( MYOD, MYOG, MRF4 ) and skeletal myocytes ( MYH3, MYH7 ) during the differentiation process. Data are mean ± SD, n = 3 independent experiments. (*P ≤ 0.05). G Immunofluorescence of MYOD, MHC, Desmin, Laminin during the differentiation of skeletal muscle cell form UiPSM at day 60 (left). The scale bar represents 100 µm. The following values represent the percentage of positive cells statistically. Data are mean ± SD, n = 3 independent experiments, each experiment counted 100 fields of view. H The UiPSM cells differentiated at day 30 and day 60 were enriched for GO terms of skeletal muscle development. I Heatmap illustrating the gene expression of skeletal muscle development related genes with dramatical change in UiPSM cell derived myocytes at day 30 and day 60

Article Snippet: Rabbit polyclonal anti-Human MYOD (Cell signaling Technology, D8G3), Mouse monoclonal anti-Myosin Heavy Chain (MF20) (R & D systems, MAB4470), Mouse monoclonal anti-Human Laminin a3 (R & D systems, MAB2144), Rabbit polyclonal anti-Human Desmin (R & D systems, abs106139), Mouse monoclonal anti-Human HNA (Millipore, MAB1281).

Techniques: In Vitro, Cell Differentiation, Gene Expression, Immunofluorescence, Derivative Assay

Journal: Cell & Bioscience

Article Title: Generation of musculoskeletal cells from human urine epithelium-derived presomitic mesoderm cells

doi: 10.1186/s13578-024-01274-w

Figure Lengend Snippet: MYOD promoted the maturity of skeletal myocytes in vitro. A Schematic overview of stepwise differentiation of skeletal myocytes from UiPSM with ectopic MYOD. SKM: skeletal muscle cells. Representative images show the morphological changes from UiPSM cells to skeletal myocytes. Scale bars, 100 µm. n = 3 independent experiments. B Representative gene expression of human skeletal muscle satellite cells (PAX3, PAX7 ), skeletal myoblasts ( MYOD, MYOG, MRF4 ) and skeletal myocytes ( MYH3, MYH7 ) when overexpressed ectopic MYOD during the differentiation process. Mcherry as a negative control of overexpression vector. Data are mean ± SD, n = 3 independent experiments. (*P ≤ 0.05). C Detection of skeletal muscle satellite cell-specific genes ( PAX3 and PAX7 ) in MYOD-mediated differentiated UiPSM cells at day 15. The following values indicate the percentage of positive cells statistically (Data are mean ± SD, n = 3 independent experiments). Scale bars, 100 µm. D Flow cytometric analysis evaluating differentiation efficiency via MHC and Desmin protein expression in skeletal muscle cells at day 60 of differentiation. hESC (H9)-derived skeletal muscle cells at day 85 are used as a positive control. E Immunofluorescence analysis of MYOD, MHC, Desmin, and Laminin in UiPSM-derived muscle fibers at day 30 with ectopic MYOD. Scale bar represents 100 µm. The values indicate the percentage of positive cells statistically (Data are mean ± SD, n = 3 independent experiments, each experiment counted 100 fields of view). Scale bars, 100 µm. F MYOD-mediated differentiation of UiPSM cells into skeletal muscle cells at days 15 and 30, showing enrichment for skeletal muscle development-related GO terms. G Heatmap illustrating gene expression changes specific to skeletal myocytes in MYOD-mediated UiPSM cell-derived myocytes at days 15 and 30

Article Snippet: Rabbit polyclonal anti-Human MYOD (Cell signaling Technology, D8G3), Mouse monoclonal anti-Myosin Heavy Chain (MF20) (R & D systems, MAB4470), Mouse monoclonal anti-Human Laminin a3 (R & D systems, MAB2144), Rabbit polyclonal anti-Human Desmin (R & D systems, abs106139), Mouse monoclonal anti-Human HNA (Millipore, MAB1281).

Techniques: In Vitro, Gene Expression, Negative Control, Over Expression, Plasmid Preparation, Expressing, Derivative Assay, Positive Control, Immunofluorescence

Journal: Cell & Bioscience

Article Title: Generation of musculoskeletal cells from human urine epithelium-derived presomitic mesoderm cells

doi: 10.1186/s13578-024-01274-w

Figure Lengend Snippet: Transplantation of UiPSM and iMYOD UiPSM cells derived human myocytes in muscle injury model. A Schematic overview of the transplantation methodology for UiPSM and iMYOD UiPSM cell-derived human skeletal myocytes into the TA muscle of MITRG mice, following treatment with cardiotoxin (CTX) for 24 h. Urine cells serve as a negative control and are transplanted into the left tibialis anterior muscle. B Bioluminescence imaging (BLI) signal captured at the right tibialis anterior graft site in a representative MITRG mouse treated with CTX, 1 month after transplantation. UCs transplanted into the left TA muscle in (a), as a negative control. UiPSM derived myocytes transplanted into the rright TA muscle in (b). C Morphological characteristics of TA muscle tissue in after transplanted UiPSM-derive myocytes and UCs. H&E staining of longitudinal sections of TA muscles showed the aggregation of inflammatory factors (asterisk) could still be seen locally in the left tibial anterior muscle after transplanting UCs in (a). H&E staining of longitudinal sections of TA muscles after transplanted UiPSM differentiated into myocytes at day 60 in (b). Scale bars, 100 µm. D Morphological characteristics of TA muscle tissue in after transplanted iMYOD UiPSM-derive myocytes and UCs. H&E staining of longitudinal sections of TA muscles showed the local aggregation of inflammatory factors (asterisk)in the left tibial anterior muscle after transplanting UCs in (a). H&E staining of longitudinal sections of TA muscles after transplanted iMYOD UiPSM-derived myocytes at day 30 in (b). Scale bars, 100 µm. E TA muscle from UiPSM-derived myocytes evaluated for the expression of myocyte-specific markers. Longitudinal section showed the colocalization of Desmin (DES) and Myosin Heavy Chain (MHC) (arrows in (a.)). Transversal section showed the colocalization of Desmin and Laminin (arrows in (b.)). Transversal section showed the colocalization of human nuclei antibody (hNA) and MYOD (arrows in (c.)). Scale bars, 100 µm. F TA muscle from iMYOD UiPSM-derived myocytes evaluated for the expression of myocyte-specific markers. Colocalization of Desmin (DES) and Myosin Heavy Chain (MHC) (arrows in (a.), and colocalization of Desmin and Laminin (arrows in (b.)) were shown in Longitudinal sections. Transversal section showed the colocalization of human nuclei antibody (hNA) and MYOD (arrows in (c.)). Scale bars, 100 µm

Article Snippet: Rabbit polyclonal anti-Human MYOD (Cell signaling Technology, D8G3), Mouse monoclonal anti-Myosin Heavy Chain (MF20) (R & D systems, MAB4470), Mouse monoclonal anti-Human Laminin a3 (R & D systems, MAB2144), Rabbit polyclonal anti-Human Desmin (R & D systems, abs106139), Mouse monoclonal anti-Human HNA (Millipore, MAB1281).

Techniques: Transplantation Assay, Derivative Assay, Negative Control, Imaging, Staining, Muscles, Expressing

Journal: iScience

Article Title: Immortalized human myotonic dystrophy type 1 muscle cell lines to address patient heterogeneity

doi: 10.1016/j.isci.2024.109930

Figure Lengend Snippet: DM1 myoblasts show higher cell proliferation and reduced levels of early myogenic markers (A) Real-time impedance curves of human iCtrl (blue) and iDM1 (red) myoblasts during culture in proliferation medium (SGM) and differentiation media (bDM and cDM). (B) Proliferation of iCtrl (blue) and iDM1 (red) myoblasts was analyzed at 24 and 48 h after seeding. (C) Differentiation of iCtrl and iDM1 myoblasts was analyzed after 2 days in differentiation medium bDM (2dpd). (D) Jess Western blot analysis of Myf5, MyoD and Myogenin in iCtrl and iDM1 3 differentiating myoblasts at 4 different time-points: 0, 1, 2 and 3 dpd. Values are represented over Ctrl 0 dpd. Data information: n = 3 for iCtrl and iDM1. Dpd, days post differentiation. All data are expressed as mean ± SEM. (A–C) Dots indicate mean values of individual samples from 10 replicates. Means were compared using unpaired two-tail Student’s t test. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01. “i” mean immortalized cell line.

Article Snippet: Rabbit polyclonal MyoD Antibody , Santa Cruz Biotechnology , Cat# sc-304, RRID: AB_631992.

Techniques: Western Blot

Journal: iScience

Article Title: Immortalized human myotonic dystrophy type 1 muscle cell lines to address patient heterogeneity

doi: 10.1016/j.isci.2024.109930

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal MyoD Antibody , Santa Cruz Biotechnology , Cat# sc-304, RRID: AB_631992.

Techniques: Virus, DNA Methylation Assay, Control, Sequencing, Software, Methylation

Journal: International Journal of Molecular Sciences

Article Title: Studying the Effect of MBNL1 and MBNL2 Loss in Skeletal Muscle Regeneration

doi: 10.3390/ijms25052687

Figure Lengend Snippet: Skeletal muscle is histologically similar between WT (FVB) and MBNL1-OE mice. ( A ) A representative image of H&E staining of TA muscles of non-injured muscle from WT and MBNL1-OE mice, and PAX7 IF (red) in TA muscle, showed increased satellite cells (white arrows) in MBNL1-OE as compared to wildtype mice. RT-PCR of Mbnl1 mRNA confirmed overexpression in MBNL1-OE mice. Muscle fiber counts showed no difference, and MBNL1-OE mice had higher counts of MuSCs in TA muscles (at least 10 independent fields from n ≥ 4 mice/group). RT-PCR showed increased Pax7 expression in MBNL1-OE mice; scale bar is 50 µm. ( B ) Immunofluorescence of recombinant MBNL1 using the anti-Flag rabbit polyclonal antibody (F7425, Sigma) in MBNL1-OE TA muscle showed MBNL1 is not expressed in the satellite cells; PAX7 (green), Flag (red), and nuclei are stained blue. Recombinant MBNL1 protein was not detected in wildtype (WT) controls. Scale bar is 10 µm. ( C ) Immunofluorescence for PAX7 (red) and MyoD (green) in non-injured TA sections from MBNL1-OE mice showed MuSCs co-expressing PAX7 and MyoD (a marker of “activated” MuSCs); this was rarely seen in WT mice; scale bar is 10 µm. ( D ) Quantitative RT-PCR of relative expression, Myog , and Myod in whole TA muscles ( n ≥ 4 mice per group) showed no significant difference. Error bars are mean ± SEM; Student’s t -test; ns = not significant; * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: Primary antibodies were anti-PAX7 (1:50, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), anti-MBNL1 (1;1000, A2764, gift from Dr. Charles A. Thornton), anti-MYH3 (1:200, clone F1.652, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), anti-laminin (1:1000, catalog L9393, Sigma-Aldrich, St. Louis, MO, USA)), anti-MyoD rabbit polyclonal (1:200, SC-760, Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Ki67 antibody rabbit polyclonal (#ab15580, Abnova, Taipei, Taiwan), and anti-Flag rabbit polyclonal (F7425, Sigma-Aldrich, St. Louis, MO, USA).

Techniques: Staining, Muscles, Reverse Transcription Polymerase Chain Reaction, Over Expression, Expressing, Immunofluorescence, Recombinant, Marker, Quantitative RT-PCR