cmv myod  (Addgene inc)

Journal: Scientific reports

Article Title: Mohawk impedes angiofibrosis by preventing the differentiation of tendon stem/progenitor cells into myofibroblasts.

doi: 10.1038/s41598-022-24195-5

Figure Lengend Snippet: Figure 3. MyoD induces TSPCs to myofibroblasts reprogramming in vitro. (A) and (B) Acta2 and Col3a1 qRT-PCR quantification comparing 3 conditions: “control TSPCs”, “MyoD-expressing TSPCs (14 days after transfection)” and “TSPCs + MyoD + Xav939 treatment”. N = 3. (C) Representative morphological change 14 days after MyoD expression under inverted (scale bar, 50 μm) or scanning electron microscope (scale bar, 25 μm). (D) PrestoBlue cell viability assay of MyoD-expressing or control TSPCs. ***p < 0.001. (E) Cellular length comparison and (F) width (Control vs. MyoD-expressing TSPCs 14 days after transfection) using image J software, 20 microscopic fields, **p < 0.01. (G) FACS gating strategy for control TSPCs and (H) myofibroblasts. (I) FACS based quantification of ACTA2 + cells. *p < 0.05. N = 3. (J) Representative immunofluorescence images of MyoD-expressing TSPCs vs. control TSPCs stained for VEGF (green) and α-SMA (red). N = 3. Scale bar, 20 μm.

Article Snippet: Neonatal TSPCs were transfected at 60–70% confluence with MyoD expressing vectors (CMV-MyoD, a gift from Andrew Lassar, Addgene plasmid # 8398) or with empty vectors (controls, CMV-MyoD restricted using EcoRI and ligated) using lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

Techniques: In Vitro, Quantitative RT-PCR, Control, Expressing, Transfection, Microscopy, Viability Assay, Comparison, Software, Immunofluorescence, Staining

Journal: Scientific reports

Article Title: Mohawk impedes angiofibrosis by preventing the differentiation of tendon stem/progenitor cells into myofibroblasts.

doi: 10.1038/s41598-022-24195-5

Figure Lengend Snippet: Figure 5. In-vivo Wnt signaling modulation during adult tendon healing. (A) qRT-PCR Acta2 and Col3a1 quantification comparing 3 conditions: (1) control TSPCs (2) MyoD-expressing TSPCs (3) TSPCs + MyoD + Xav939 treatment. N = 3. (B) Representative H&E and Masson’s trichrome staining images of control and Xav939-treated tendons d28 after injury. N = 6 tendons/group. Scale bar, 40 μm, and (C) the corresponding Collagen deposition quantification. (D) Width quantification of control and Xav939-treated Achilles tendons (20 zones along 5 tendons/group). (E) Tendon thickness of control and Xav939-treated tendons (20 zones from 5 tendons/group). (F–I) Maximum force, yield elongation, stiffness and maximum tensile stress properties in the control and Xav939-treated groups d52 after injury. N = 6. Representative immunofluorescence staining images of α-SMA and VEGF.N = 5 tendons/group. Scale bar, 40 μm.

Article Snippet: Neonatal TSPCs were transfected at 60–70% confluence with MyoD expressing vectors (CMV-MyoD, a gift from Andrew Lassar, Addgene plasmid # 8398) or with empty vectors (controls, CMV-MyoD restricted using EcoRI and ligated) using lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

Techniques: In Vivo, Quantitative RT-PCR, Control, Expressing, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Mohawk impedes angiofibrosis by preventing the differentiation of tendon stem/progenitor cells into myofibroblasts

doi: 10.1038/s41598-022-24195-5

Figure Lengend Snippet: MyoD induces TSPCs to myofibroblasts reprogramming in vitro. ( A ) and (B ) Acta2 and Col3a1 qRT-PCR quantification comparing 3 conditions: “control TSPCs”, “MyoD-expressing TSPCs (14 days after transfection)” and “TSPCs + MyoD + Xav939 treatment”. N = 3. ( C ) Representative morphological change 14 days after MyoD expression under inverted (scale bar, 50 μm) or scanning electron microscope (scale bar, 25 μm). ( D ) PrestoBlue cell viability assay of MyoD -expressing or control TSPCs. ***p < 0.001. ( E ) Cellular length comparison and ( F ) width (Control vs. MyoD-expressing TSPCs 14 days after transfection) using image J software, 20 microscopic fields, **p < 0.01. ( G ) FACS gating strategy for control TSPCs and ( H ) myofibroblasts. ( I ) FACS based quantification of ACTA2 + cells. *p < 0.05. N = 3. ( J ) Representative immunofluorescence images of MyoD-expressing TSPCs vs . control TSPCs stained for VEGF (green) and α-SMA (red). N = 3. Scale bar, 20 μm.

Article Snippet: Neonatal TSPCs were transfected at 60–70% confluence with MyoD expressing vectors (CMV- MyoD , a gift from Andrew Lassar, Addgene plasmid # 8398) or with empty vectors (controls, CMV- MyoD restricted using EcoRI and ligated) using lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

Techniques: In Vitro, Quantitative RT-PCR, Control, Expressing, Transfection, Microscopy, Viability Assay, Comparison, Software, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Mohawk impedes angiofibrosis by preventing the differentiation of tendon stem/progenitor cells into myofibroblasts

doi: 10.1038/s41598-022-24195-5

Figure Lengend Snippet: In-vivo Wnt signaling modulation during adult tendon healing. ( A ) qRT-PCR Acta2 and Col3a1 quantification comparing 3 conditions: (1) control TSPCs (2) MyoD-expressing TSPCs (3) TSPCs + MyoD + Xav939 treatment. N = 3. ( B ) Representative H&E and Masson’s trichrome staining images of control and Xav939-treated tendons d28 after injury. N = 6 tendons/group. Scale bar, 40 μm, and ( C ) the corresponding Collagen deposition quantification . ( D ) Width quantification of control and Xav939-treated Achilles tendons (20 zones along 5 tendons/group). ( E ) Tendon thickness of control and Xav939-treated tendons (20 zones from 5 tendons/group). ( F–I ) Maximum force, yield elongation, stiffness and maximum tensile stress properties in the control and Xav939-treated groups d52 after injury. N = 6. Representative immunofluorescence staining images of α-SMA and VEGF.N = 5 tendons/group. Scale bar, 40 μm.

Article Snippet: Neonatal TSPCs were transfected at 60–70% confluence with MyoD expressing vectors (CMV- MyoD , a gift from Andrew Lassar, Addgene plasmid # 8398) or with empty vectors (controls, CMV- MyoD restricted using EcoRI and ligated) using lipofectamine 2000 reagent (Invitrogen, USA) according to the manufacturer’s recommendations.

Techniques: In Vivo, Quantitative RT-PCR, Control, Expressing, Staining, Immunofluorescence

Journal: Stem cells (Dayton, Ohio)

Article Title: Regulation of Myogenesis by a Na/K-ATPase α1 Caveolin Binding Motif

doi: 10.1093/stmcls/sxab012

Figure Lengend Snippet: Primer sequences

Article Snippet: The primer sequences are listed in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Primer Name Primer Sequence (5'→3') human HPRT1 Forward TGGACAGGACTGAACGTCTT human HPRT1 Reverse TCCAGCAGGTCAGCAAAGAA Human MYOD Forward CGACGGCATGATGGACTACA Human MYOD Reverse GGCAGTCTAGGCTCGACAC Human MYOG Forward CCAGGGGATCATCTGCTCACG Human MYOG Reverse GGAAGGCCACAGACACATCT Human MYH8 Forward GCAGACAGAAGCGGGTGAAT Human MYH8 Reverse GCTGAGTAGATGCTTGCTTGC Human MYH3 Forward GAGGCTGGTGAGCTGAGTC Human MYH3 Reverse GCTGCCTCTTGAGCTCTTCT Human TNNT1 Forward ACCTGGTCAAGGCAGAACAG Human TNNT1 Reverse TGTCCAGAGGCTTCTTACGC Human CAV3 Forward ACCTTCTGCAACCCACTCTTC Human CAV3 Reverse GAGCAGTCCCTGGCTTTAGAC Human MYF5 Forward TGAGAGAGCAGGTGGAGAACT Human MYF5 Reverse ACATTCGGGCATGCCATCAGA Human MSGN1 Forward TGTTGGACCCACCAGAACAC Human MSGN1 reverse TTGCAAAGGATGAGCCTCCC Human PAX3 Forward CAAGCCCAAGCAGGTGACAA Human PAX3 reverse TCGGATTTCCCAGCTGAACA Human MIXL1 Forward AGTCCAGGATCCAGGTATGGT Human MIXL1 Reverse GGCCTAGCCAAAGGTTGGAA Human CTNNB1 Forward GGCTACTCAAGCTGATTTGATGG Human CTNNB1 Reverse AAGACTGTTGCTGCCAGTGA Human TBXT Forward GGGTACTCCCAATCCTATTCTGAC Human TBXT Reverse ACTGACTGGAGCTGGTAGGT Human NANOG Forward CAATGGTGTGACGCAGGGAT Human NANOG Reverse TGCACCAGGTCTGAGTGTTC Human OCT4 Forward CGAGAAGGATGTGGTCCGAG Human OCT4 Reverse GAGACAGGGGGAAAGGCTTC Human PPARG Forward GATACACTGTCTGCAAACATATCACAA Human PPARG Reverse CCACGGAGCTGATCCCAA Human FASN Forward TCGTGGGCTACAGCATGGT Human FASN Reverse GCCCTCTGAAGTCGAAGAAGAA Human ADIPOQ Forward TCTGCCTTCCGCAGTGTAGG Human ADIPOQ Reverse GGTGTGGCTTGGGGATACGA Human FABP4 Forward GCTTTGCCACCAGGAAAGTG Human FABP4 Reverse ATGGACGCATTCCACCACCA Mouse Myog Forward GTCCCAACCCAGGAGATCATT Mouse Myog Reverse AGTTGGGCATGGTTTCGTCT Mouse Myod1 Forward TGGCATGATGGATTACAGCGG Mouse Myod1 Reverse GGTGGTGCATCTGCCAAAAG mouse Myh3 Forward CTCTGTCACAGTCAGAGGTGT mouse Myh3 Reverse CTTTTCCGACTTGCGGAGGA mouse Myh8 Forward CTGAGGAGGCTGAGGAACAATC mouse Myh8 Reverse CCTCCTTCCTCTGCAAGATGT Mouse Actb Forward GGCTGTATTCCCCTCCATCG Mouse Actb Reverse CCAGTTGGTAACAATGCCATGT Open in a separate window Primer sequences Lentivirus-mediated gene delivery Lentivirus expression vector for the mouse Myod1 gene (pLv-CMV-Myod) was from Addgene (Plasmid #26808).

Techniques: Sequencing

Journal: Stem cells (Dayton, Ohio)

Article Title: Regulation of Myogenesis by a Na/K-ATPase α1 Caveolin Binding Motif

doi: 10.1093/stmcls/sxab012

Figure Lengend Snippet: Rescue of Skm differentiation in mCBM cells with a mouse Myod1 transgene.

Article Snippet: The primer sequences are listed in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Primer Name Primer Sequence (5'→3') human HPRT1 Forward TGGACAGGACTGAACGTCTT human HPRT1 Reverse TCCAGCAGGTCAGCAAAGAA Human MYOD Forward CGACGGCATGATGGACTACA Human MYOD Reverse GGCAGTCTAGGCTCGACAC Human MYOG Forward CCAGGGGATCATCTGCTCACG Human MYOG Reverse GGAAGGCCACAGACACATCT Human MYH8 Forward GCAGACAGAAGCGGGTGAAT Human MYH8 Reverse GCTGAGTAGATGCTTGCTTGC Human MYH3 Forward GAGGCTGGTGAGCTGAGTC Human MYH3 Reverse GCTGCCTCTTGAGCTCTTCT Human TNNT1 Forward ACCTGGTCAAGGCAGAACAG Human TNNT1 Reverse TGTCCAGAGGCTTCTTACGC Human CAV3 Forward ACCTTCTGCAACCCACTCTTC Human CAV3 Reverse GAGCAGTCCCTGGCTTTAGAC Human MYF5 Forward TGAGAGAGCAGGTGGAGAACT Human MYF5 Reverse ACATTCGGGCATGCCATCAGA Human MSGN1 Forward TGTTGGACCCACCAGAACAC Human MSGN1 reverse TTGCAAAGGATGAGCCTCCC Human PAX3 Forward CAAGCCCAAGCAGGTGACAA Human PAX3 reverse TCGGATTTCCCAGCTGAACA Human MIXL1 Forward AGTCCAGGATCCAGGTATGGT Human MIXL1 Reverse GGCCTAGCCAAAGGTTGGAA Human CTNNB1 Forward GGCTACTCAAGCTGATTTGATGG Human CTNNB1 Reverse AAGACTGTTGCTGCCAGTGA Human TBXT Forward GGGTACTCCCAATCCTATTCTGAC Human TBXT Reverse ACTGACTGGAGCTGGTAGGT Human NANOG Forward CAATGGTGTGACGCAGGGAT Human NANOG Reverse TGCACCAGGTCTGAGTGTTC Human OCT4 Forward CGAGAAGGATGTGGTCCGAG Human OCT4 Reverse GAGACAGGGGGAAAGGCTTC Human PPARG Forward GATACACTGTCTGCAAACATATCACAA Human PPARG Reverse CCACGGAGCTGATCCCAA Human FASN Forward TCGTGGGCTACAGCATGGT Human FASN Reverse GCCCTCTGAAGTCGAAGAAGAA Human ADIPOQ Forward TCTGCCTTCCGCAGTGTAGG Human ADIPOQ Reverse GGTGTGGCTTGGGGATACGA Human FABP4 Forward GCTTTGCCACCAGGAAAGTG Human FABP4 Reverse ATGGACGCATTCCACCACCA Mouse Myog Forward GTCCCAACCCAGGAGATCATT Mouse Myog Reverse AGTTGGGCATGGTTTCGTCT Mouse Myod1 Forward TGGCATGATGGATTACAGCGG Mouse Myod1 Reverse GGTGGTGCATCTGCCAAAAG mouse Myh3 Forward CTCTGTCACAGTCAGAGGTGT mouse Myh3 Reverse CTTTTCCGACTTGCGGAGGA mouse Myh8 Forward CTGAGGAGGCTGAGGAACAATC mouse Myh8 Reverse CCTCCTTCCTCTGCAAGATGT Mouse Actb Forward GGCTGTATTCCCCTCCATCG Mouse Actb Reverse CCAGTTGGTAACAATGCCATGT Open in a separate window Primer sequences Lentivirus-mediated gene delivery Lentivirus expression vector for the mouse Myod1 gene (pLv-CMV-Myod) was from Addgene (Plasmid #26808).

Techniques: