Journal: Biomaterials Research

Article Title: Chicken Embryo Extract Remodeling of Extracellular Matrix Sustains Self-Renewal and Differentiation for Scaffold-Free Cell Sheet Formation

doi: 10.34133/bmr.0332

Figure Lengend Snippet: Expression of myogenic transcription factors in PMSCs according to PM composition. (A) Immunofluorescent staining results for PAX7 (green), MYOD (red), and DAPI (blue). (B) Immunofluorescent staining results for Myogenin (green) and DAPI (blue). (C to E) Percentages of PAX7, MYOD1, and Myogenin-positive cells. All percentages were determined by counting PAX7, MYOD, and Myogenin-positive cells in relation to the total number of DAPI-positive cells. (F) Protein expression levels of PAX7, MYOD1, Myogenin, and GAPDH in PMSCs under different culture conditions. (G) PAX7, MYOD, and Myogenin protein expression levels were normalized to GAPDH levels. (H) mRNA expression levels of PAX7 , MYF5 , MYOD1 , and Myogenin in PMSCs. n = 3. All values are represented as means ± SE. a-c Different superscripts represent statistically significant differences ( P < 0.01). A-B Different superscripts represent statistically significant differences ( P < 0.05). ns, nonsignificant.

Article Snippet: Each membrane was incubated overnight at 4 °C with the following antibodies: PAX7 (1:50; DSHB), MYOD1 (1:1,000; Proteintech), Myogenin (1:50; DSHB), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1: 5,000, Invitrogen).

Techniques: Expressing, Staining

Journal: bioRxiv

Article Title: NRMLncR, a myocyte-enriched long non-coding RNA, enhances myogenesis in mouse

doi: 10.64898/2026.02.11.704964

Figure Lengend Snippet: (A) Coding Potential Assessment Tool (CPAT) analysis showing low protein-coding potential of NRMLncR-v1 and NRMLncR-v2 . Coding probability >=0.44 indicates coding protein. (B) Western blot analysis of FLAG tag to detect predicted proteins encoded by NRMLncR-v1 and NRMLncR-v2 open reading frames and their mutant ORF. FLAG-tagged HEY1 served as a positive control. (C and D) Immunofluorescence staining showing no detectable protein expression from the predicted NRMLncR-v1 (C) and NRMLncR-v2 (D) open reading frames in HEK293 cells, compared with a FLAG-tagged HEY1 positive control. Scale bar: 50 μm. (E) Luciferase reporter assays showing mildly inhibition of the NRMLncR promoter activity by RBPJ. n=4. (F) Schematic representation of putative E-box motifs within the NRMLncR promoter region. (G) ChIP-PCR and DNA gel analysis indicating the binding of MyoD to the E-box sites on the NRMLncR promoter. (H) ChIP-qPCR analysis showing enrichment of MyoD at the E-box regions of the NRMLncR promoter. n=3. (I) NRMLncR-v1/2 expression levels in myoblasts overexpressing MyoD. n=4. (J) Luciferase reporter assays showing transcriptional activation of the NRMLncR promoter by MyoD, MyoG, and MRF4. n=4. Data represent mean ± SEM. One-way ANOVA multiple comparisons test (E, H, J), Student’s t -test (I): * p < .05; ** p < .01; and *** p < .001.

Article Snippet: Chromatin was immunoprecipitated using antibodies against MyoD (Santa Cruz Biotechnology, cat#sc-377460) and MyoG (DSHB, cat#F5D), with mouse IgG (Santa Cruz Biotechnology, cat#sc-2025) serving as a negative control.

Techniques: Western Blot, FLAG-tag, Mutagenesis, Positive Control, Immunofluorescence, Staining, Expressing, Luciferase, Inhibition, Activity Assay, Binding Assay, ChIP-qPCR, Activation Assay

Journal: eLife

Article Title: Branched actin polymerization drives invasive protrusion formation to promote myoblast fusion during mouse skeletal muscle regeneration

doi: 10.7554/eLife.103550

Figure Lengend Snippet: ( A ) Schematic diagram of tamoxifen and BaCI 2 treatment and subsequent cell proliferation and differentiation analyses at dpi 2.5 and 4, respectively. ( B ) Immunostaining with anti-Laminin, anti-Pax7, and anti-Ki67 of cross-sections of TA muscles from Ctrl and mutant mice at dpi 2.5. The boxed areas are shown on the right. Scale bar: 30 µm. ( C ) Immunostaining with anti-Laminin and anti-MyoG of cross-sections of TA muscles from Ctrl and mutant mice at dpi 4. The boxed areas are shown at the top-right corner. Scale bar: 30 µm. ( D ) Quantification of the percentage of proliferating satellite cells (% of Ki67 + cells in the Pax7 + cells) in TA muscles from Ctrl and mutant mice of the indicated genotypes. n=3 mice of each genotype were examined. Mean ± s.d. values are shown in the bar graph, and significance was determined by two-tailed Student’s t -test. ns: not significant. ( E ) Quantification of the percentage of MyoG + nuclei in the total cells in TA muscles from Ctrl and mutant mice of the indicated genotypes. n=3 mice of each genotype were examined. Mean ± s.d. values are shown in the bar graph, and significance was determined by two-tailed Student’s t -test. ns: not significant. ( F ) Western blot analysis for MymX and MymK in TA muscles from Ctrl and mutant mice of the indicated genotypes at dpi 4. One sample of each Ctrl and mutant genotype is shown. ( G, H ) Quantification of MymX and MymK protein expression in the Ctrl and littermate mutant mice as shown in ( F ). The band intensity of each protein was normalized against β-tubulin. The y axis indicates the expression of MymX or MymK in different mutants relative to the control mice. n=3 mice of each genotype were examined. Mean ± s.d. values are shown in the bar graphs, and significance was determined by two-tailed Student’s t -test. *p<0.05; ns: not significant. Figure 3—figure supplement 1—source data 1. PDF file containing original western blots for , indicating the relevant bands and treatments. Figure 3—figure supplement 1—source data 2. Original files for western blot analysis displayed in .

Article Snippet: Then, the sections were fixed in 2% PFA for 5 minutes at RT, washed three times with PBS, and incubated with blocking buffer supplemented with M.O.M blocking reagent (1:25; Vector; MKB-2213-1) for 60 minutes at RT, followed by overnight incubation with mouse anti-Pax7 (1:2; DSHB; Pax7), mouse anti-MyoG (1:2; DSHB; F5D), rat anti-Laminin-2 (1:500; Sigma; L0663), and rat anti-Ki67 (1:500; Thermo Fisher Scientific; 14-5698-82) at 4°C.

Techniques: Immunostaining, Muscles, Mutagenesis, Two Tailed Test, Western Blot, Expressing, Control