Journal: Frontiers in Genetics
Article Title: Effects and Molecular Mechanism of Single-Nucleotide Polymorphisms of MEG3 on Porcine Skeletal Muscle Development
doi: 10.3389/fgene.2021.607910
Figure Lengend Snippet: Identification of SCs, construction of the expression vector and the effect of two haplotypes on MEG3 transcription. (A) SCs were identified by immunolabeling with anti-Pax7 (red) monolyclonal antibody; Nuclear are counterstained with DAPI (blue); Merged images represented overlays of Pax7 (red) and nuclear staining by DAPI (blue). (B) A FLAG tag sequence (pink) was added at the 3′ end of the MEG3 sequence (gray). This fragment was inserted into the pcDNA3.1(+) vector between the EcoR I and Xho I site. (C) The RT-PCR results of SCs transfected with pcDNA3.1-MEG3-CCCA, pcDNA3.1-MEG3-TTCC and pcDNA 3.1 (+) vector for 48 h. M: Marker 2000, Line 1–3: the RT-PCR results of pcDNA3.1-MEG3-CCCA group, pcDNA3.1-MEG3-TTCC group and pcDNA 3.1(+) group were amplified by MEG3-FLAG, respectively, Line 4: the RT-PCR results of pcDNA 3.1(+) group were amplified by Primer-GAPDH. (D) The effect of two haplotypes on MEG3 transcription. The significance test: ∗∗ P < 0.01.
Article Snippet: The isolated SCs were seeded in 24-well plates and identified by immunofluorescence staining with anti-mouse Paired box protein Pax7 monoclonal antibody (Santa Cruz Biotechnology, Inc., Europe).
Techniques: Expressing, Plasmid Preparation, Immunolabeling, Staining, FLAG-tag, Sequencing, Reverse Transcription Polymerase Chain Reaction, Transfection, Marker, Amplification