Review



phosphorylated myod  (Santa Cruz Biotechnology)


Bioz Verified Symbol Santa Cruz Biotechnology is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 95
      Buy from Supplier

    Structured Review

    Santa Cruz Biotechnology phosphorylated myod
    Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of <t>MyoD,</t> myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.
    Phosphorylated Myod, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 670 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+myod/MyoD/pm37174683-62-5-8
    Average 95 stars, based on 670 article reviews
    phosphorylated myod - by Bioz Stars, 2026-09
    95/100 stars

    Images

    1) Product Images from "IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger."

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.

    Journal: Cells

    doi: 10.3390/cells12091284

    Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of MyoD, myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.
    Figure Legend Snippet: Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of MyoD, myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.

    Techniques Used: Expressing, Incubation, Quantitative RT-PCR

    Figure 3. Reduction of IL-4/IL-4Rα signaling suppressed the increased expression of MyoD, myo- genin, and myomerger, but did not affect myomaker expression. (A) Representative immunofluores- cence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. Cell nuclei were counterstained with DAPI (blue). C2C12 cells transfected with control siRNA (Ctrl) or IL-4Rα siRNA (IL-4RαKD) were grown in GM for 24 h and maintained in DM with or without recombinant IL-4 (10 ng/mL) for 72 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells was significantly sup- pressed by IL-4Rα knockdown. N = 5 per group. (C–E) Reduction of IL-4/IL-4Rα signaling by IL-4Rα knockdown significantly suppressed the increased expression of MyoD, myogenin, and my- omerger, whereas the expression of myomaker was not affected. Cells were treated as described in (A). (C,E) show fold changes in mRNA and protein levels, respectively. N = 6 per group. Representative western blot analysis is shown in (D). Data represent mean ± SD. * p < 0.05.
    Figure Legend Snippet: Figure 3. Reduction of IL-4/IL-4Rα signaling suppressed the increased expression of MyoD, myo- genin, and myomerger, but did not affect myomaker expression. (A) Representative immunofluores- cence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. Cell nuclei were counterstained with DAPI (blue). C2C12 cells transfected with control siRNA (Ctrl) or IL-4Rα siRNA (IL-4RαKD) were grown in GM for 24 h and maintained in DM with or without recombinant IL-4 (10 ng/mL) for 72 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells was significantly sup- pressed by IL-4Rα knockdown. N = 5 per group. (C–E) Reduction of IL-4/IL-4Rα signaling by IL-4Rα knockdown significantly suppressed the increased expression of MyoD, myogenin, and my- omerger, whereas the expression of myomaker was not affected. Cells were treated as described in (A). (C,E) show fold changes in mRNA and protein levels, respectively. N = 6 per group. Representative western blot analysis is shown in (D). Data represent mean ± SD. * p < 0.05.

    Techniques Used: Expressing, Transfection, Control, Recombinant, Knockdown, Western Blot

    Figure 4. Stimulation of IL-4/IL-4Rα signaling increased the expression of myogenic regulatory factors and myomerger, but not myomaker, even in GM. (A) Representative immunofluorescence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. The nuclei were counterstained with DAPI (blue). C2C12 cells were treated as in Figure 3 except that cells were maintained in GM, instead of DM, for 24 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells significantly increased by IL-4 treatment, which was suppressed by IL-4Rα knockdown. N = 4 per group. (C) Fold change in the mRNA levels of MyoD, myogenin, myomerger, and myomaker. N = 6 per group. Data represent as mean ± SD. * p < 0.05.
    Figure Legend Snippet: Figure 4. Stimulation of IL-4/IL-4Rα signaling increased the expression of myogenic regulatory factors and myomerger, but not myomaker, even in GM. (A) Representative immunofluorescence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. The nuclei were counterstained with DAPI (blue). C2C12 cells were treated as in Figure 3 except that cells were maintained in GM, instead of DM, for 24 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells significantly increased by IL-4 treatment, which was suppressed by IL-4Rα knockdown. N = 4 per group. (C) Fold change in the mRNA levels of MyoD, myogenin, myomerger, and myomaker. N = 6 per group. Data represent as mean ± SD. * p < 0.05.

    Techniques Used: Expressing, Knockdown

    Related Articles

    Expressing:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Incubation:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Quantitative RT-PCR:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Transfection:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Control:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Recombinant:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Knockdown:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).

    Western Blot:

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.
    Article Snippet: Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].Cultured C2C12 cells were homogenized in ice-cold radioimmunoprecipitation assay (RIPA) buffer (Wako) with protease and phosphatase inhibitors (Nacalai Tesque Inc.) and processed for western blotting analyses as previously described [25,26].. Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).. Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).Luminescence signals from ECL reagent (Bio-Rad Laboratories Inc., Hercules, CA, USA) were detected using LAS-4000 (Fujifilm Corp., Tokyo, Japan).



    Similar Products

    phosphorylated myod  (Santa Cruz Biotechnology)
    95
    Santa Cruz Biotechnology phosphorylated myod
    Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of <t>MyoD,</t> myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.
    Phosphorylated Myod, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+myod/MyoD/pm37174683-62-5-8
    Average 95 stars, based on 1 article reviews
    phosphorylated myod - by Bioz Stars, 2026-09
    95/100 stars
    		  Buy from Supplier
    mos phosphorylation of myod-ser237  (Federation of European Neuroscience Societies)
    90
    Federation of European Neuroscience Societies mos phosphorylation of myod-ser237
    Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of <t>MyoD,</t> myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.
    Mos Phosphorylation Of Myod Ser237, supplied by Federation of European Neuroscience Societies, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/phosphorylated+myod/mos+phosphorylation+of+myod+ser237/pm10838091-4-9-25
    Average 90 stars, based on 1 article reviews
    mos phosphorylation of myod-ser237 - by Bioz Stars, 2026-09
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of MyoD, myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.

    Journal: Cells

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.

    doi: 10.3390/cells12091284

    Figure Lengend Snippet: Figure 1. IL-4 treatment promoted myoblast differentiation and increased the expression of myogenic regulatory factors and myomerger in C2C12 cells. (A) Representative immunofluorescence images of MyHC (red) in the differentiated C2C12 cells 72 h after IL-4 application. The nuclei of C2C12 cells were counterstained with DAPI (blue). Scale bars: 50 µm. (B,C) The number of myonuclei per myotube and the differentiation index of IL-4-treated C2C12 cells. N = 3 per group. (D) Representative images of EdU (green) and counterstained nuclei (Hoechst 33342; blue) after 24 h IL-4 treatment in GM. (E) The number of EdU-positive cells significantly decreased in the IL-4-treated C2C12 cells. N = 4 per group. (F) Fold changes in the mRNA levels of MyoD, myogenin, myomaker, and myomerger in the IL-4-treated C2C12 myoblasts. Myoblasts were maintained in DM with 10 ng/mL IL-4. After 72 h incubation, the cells were harvested for qRT-PCR. N = 5 per group. Data are presented as mean ± SD. * p < 0.05.

    Article Snippet: Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).

    Techniques: Expressing, Incubation, Quantitative RT-PCR

    Figure 3. Reduction of IL-4/IL-4Rα signaling suppressed the increased expression of MyoD, myo- genin, and myomerger, but did not affect myomaker expression. (A) Representative immunofluores- cence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. Cell nuclei were counterstained with DAPI (blue). C2C12 cells transfected with control siRNA (Ctrl) or IL-4Rα siRNA (IL-4RαKD) were grown in GM for 24 h and maintained in DM with or without recombinant IL-4 (10 ng/mL) for 72 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells was significantly sup- pressed by IL-4Rα knockdown. N = 5 per group. (C–E) Reduction of IL-4/IL-4Rα signaling by IL-4Rα knockdown significantly suppressed the increased expression of MyoD, myogenin, and my- omerger, whereas the expression of myomaker was not affected. Cells were treated as described in (A). (C,E) show fold changes in mRNA and protein levels, respectively. N = 6 per group. Representative western blot analysis is shown in (D). Data represent mean ± SD. * p < 0.05.

    Journal: Cells

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.

    doi: 10.3390/cells12091284

    Figure Lengend Snippet: Figure 3. Reduction of IL-4/IL-4Rα signaling suppressed the increased expression of MyoD, myo- genin, and myomerger, but did not affect myomaker expression. (A) Representative immunofluores- cence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. Cell nuclei were counterstained with DAPI (blue). C2C12 cells transfected with control siRNA (Ctrl) or IL-4Rα siRNA (IL-4RαKD) were grown in GM for 24 h and maintained in DM with or without recombinant IL-4 (10 ng/mL) for 72 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells was significantly sup- pressed by IL-4Rα knockdown. N = 5 per group. (C–E) Reduction of IL-4/IL-4Rα signaling by IL-4Rα knockdown significantly suppressed the increased expression of MyoD, myogenin, and my- omerger, whereas the expression of myomaker was not affected. Cells were treated as described in (A). (C,E) show fold changes in mRNA and protein levels, respectively. N = 6 per group. Representative western blot analysis is shown in (D). Data represent mean ± SD. * p < 0.05.

    Article Snippet: Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).

    Techniques: Expressing, Transfection, Control, Recombinant, Knockdown, Western Blot

    Figure 4. Stimulation of IL-4/IL-4Rα signaling increased the expression of myogenic regulatory factors and myomerger, but not myomaker, even in GM. (A) Representative immunofluorescence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. The nuclei were counterstained with DAPI (blue). C2C12 cells were treated as in Figure 3 except that cells were maintained in GM, instead of DM, for 24 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells significantly increased by IL-4 treatment, which was suppressed by IL-4Rα knockdown. N = 4 per group. (C) Fold change in the mRNA levels of MyoD, myogenin, myomerger, and myomaker. N = 6 per group. Data represent as mean ± SD. * p < 0.05.

    Journal: Cells

    Article Title: IL-4 Signaling Promotes Myoblast Differentiation and Fusion by Enhancing the Expression of MyoD, Myogenin, and Myomerger.

    doi: 10.3390/cells12091284

    Figure Lengend Snippet: Figure 4. Stimulation of IL-4/IL-4Rα signaling increased the expression of myogenic regulatory factors and myomerger, but not myomaker, even in GM. (A) Representative immunofluorescence images of myogenin (red) in the IL-4RαKD C2C12 myoblasts. The nuclei were counterstained with DAPI (blue). C2C12 cells were treated as in Figure 3 except that cells were maintained in GM, instead of DM, for 24 h. Scale bars: 50 µm. (B) The percentage of myogenin-positive cells significantly increased by IL-4 treatment, which was suppressed by IL-4Rα knockdown. N = 4 per group. (C) Fold change in the mRNA levels of MyoD, myogenin, myomerger, and myomaker. N = 6 per group. Data represent as mean ± SD. * p < 0.05.

    Article Snippet: Primary antibodies used were against phosphorylated MyoD (1:1000; Santa Cruz Biotechnology Inc.), myogenin (1:1000; DSHB), myomaker (1:1000, Abcam, Cambridge, UK), myomerger (1:1000; R&D Systems), IL-4Rα (1:1000; Santa Cruz Biotechnology Inc.), MyHC (1:200; DSHB), and GAPDH (1:2000; Cell Signaling Technology Inc., Danvers, MA, USA).

    Techniques: Expressing, Knockdown