Journal: International Journal of Molecular Sciences

Article Title: Human Wharton’s Jelly-Derived Mesenchymal Stem Cells Minimally Improve the Growth Kinetics and Cardiomyocyte Differentiation of Aged Murine Cardiac c-kit Cells in In Vitro without Rejuvenating Effect

doi: 10.3390/ijms20225519

Figure Lengend Snippet: List of antibodies for CC characterisation and differentiation.

Article Snippet: FITC Rat Anti-mouse Sca-1 Antibody (Clone D7) , 1:10 , FC , Miltenyi Biotec, Germany (130-102-297).

Techniques:

Journal: eLife

Article Title: Epigenetic regulation of Wnt7b expression by the cis -acting long noncoding RNA Lnc-Rewind in muscle stem cells

doi: 10.7554/eLife.54782

Figure Lengend Snippet: ( A ) Table represents the values obtained by analysing the local sequence alignment between the human and murine Lnc-Rewind transcripts. Data were produced by using the implementation of the Smith–Waterman algorithm available at http://www.ebi.ac.uk/Tools/psa/emboss_water/ . ( B ) Semiquantitative RT-PCR (sqRT-PCR) quantification of the human hs_Lnc-Rewind transcript in proliferating (GM) myoblasts from a healthy donor. Mature Gapdh was used as endogenous control. ( C ) FACS plot showing MuSC isolation strategy from WT mice. MuSCs are isolated, among the lineage (CD45/CD31/Ter119) negative cells, as α7integrin+/Sca1− cells (magenta box) (left and middle panels). The right plot shows the check purity of sorted MuSCs. ( D ) qRT-PCR quantification of Myog and Mck in C 2 C 12 and MuSC-derived myoblasts in growth (GM) and differentiated (DM) conditions. Data represent the mean ± SEM of three biological replicates and were normalized on Gapdh mRNA. ( E ) sqRT-PCR quantification of Lnc-Rewind, using different primers, in cytoplasmic (Cyt) and nuclear (Nuc) fractions from proliferating C 2 C 12 and MuSC-derived myoblasts. The quality of fractionation was tested with mature ( Gapdh ) and precursor ( pre-Gapdh ) RNAs. –rt represents the negative control ( F ) Representative 60X confocal images of MuSC-derived myoblasts cell cultures hybridized with probes set specific for Lnc-Rewind (upper panels) and for a human mRNA (dlc1), as negative control (neg ctrl) (bottom panels). Autofluorescence (gray) is shown with false colour to visualize the cell body. DAPI, 4’,6-diamidino-2-phenylindole (blue); scale bar: 25 μm. ( G ) Representative 60× confocal images of C 2 C 12 cell cultures hybridized with probes set specific for Lnc-Rewind (upper panels) and for a human mRNA (dlc1), as negative control (neg ctrl) (bottom panels). Autofluorescence (grey) is shown with false colour to visualize the cell body. DAPI, 4’,6-diamidino-2-phenylindole (blue); scale bar: 25 μm. Data information: *p<0.05, unpaired Student’s t-test. Figure 1—figure supplement 1—source data 1. Source data for .

Article Snippet: Cells were incubated with primary antibodies CD31-PE (MiltenyBiotec, 130111540; RRID: AB_2657296 ), CD45-PE (MiltenyBiotec, 139110797; RRID: AB_2658218 ), Ter119-PE (MiltenyBiotec, 130112909; RRID: AB_2654115 ) 1:25; Sca1-FITC (MiltenyBiotec, 130116490; RRID: AB_2751322 ) 1:50; α7Integrin-APCVio770 (MiltenyBiotec, 130095212; Custom) 1:20 for 45 min on ice diluted in HBSS containing 0.2% BSA, 1% penicillin-streptomycin, and 1% DNAse I.

Techniques: Sequencing, Produced, Reverse Transcription Polymerase Chain Reaction, Control, Isolation, Quantitative RT-PCR, Derivative Assay, Fractionation, Negative Control

Journal: eLife

Article Title: Epigenetic regulation of Wnt7b expression by the cis -acting long noncoding RNA Lnc-Rewind in muscle stem cells

doi: 10.7554/eLife.54782

Figure Lengend Snippet:

Article Snippet: Cells were incubated with primary antibodies CD31-PE (MiltenyBiotec, 130111540; RRID: AB_2657296 ), CD45-PE (MiltenyBiotec, 139110797; RRID: AB_2658218 ), Ter119-PE (MiltenyBiotec, 130112909; RRID: AB_2654115 ) 1:25; Sca1-FITC (MiltenyBiotec, 130116490; RRID: AB_2751322 ) 1:50; α7Integrin-APCVio770 (MiltenyBiotec, 130095212; Custom) 1:20 for 45 min on ice diluted in HBSS containing 0.2% BSA, 1% penicillin-streptomycin, and 1% DNAse I.

Techniques: Sequencing, Control, Clone Assay, SYBR Green Assay, Magnetic Beads, Recombinant, Software

Journal: eLife

Article Title: Tgfβ signaling is critical for maintenance of the tendon cell fate

doi: 10.7554/eLife.52695

Figure Lengend Snippet: ( A ) The colony-forming efficiency of P7 wild-type and heterozygous tenocytes as well as mutant tendon cells were evaluated by seeding one cell per well of the FACS-sorted cells in 96-well plates, and colonies formed were visualized with crystal violet staining. Mutant tenocytes exhibited significantly higher clonogenic capacity compared to wild-type and heterozygous controls. The results shown are mean ± SD (n = 5–6, **p<0.01). ( B ) Immunofluorescence staining for stem/progenitor markers in transverse sections of mutant tendons shows that mutant tendon cells acquired in postnatal stages expression of stem cell antigen-1 (Sca-1) and CD44. ( C ) In wild-type littermate controls, expression of both markers was detected in epitenon (white arrowhead), but not in tenocytes. Dashed line demarcates the mutant tendon. Scale bars, 10 μm. Mutant: CKO, Wild-type: WT, Heterozygous: Het.

Article Snippet: Antibody , Goat anti-Sca-1/Ly6 , R and D Systems , Cat# AF1226 RRID: AB_354679 , IF(1:80).

Techniques: Mutagenesis, Staining, Immunofluorescence, Expressing

Journal: eLife

Article Title: Tgfβ signaling is critical for maintenance of the tendon cell fate

doi: 10.7554/eLife.52695

Figure Lengend Snippet: ( A–D ) Immunofluorescence staining for Sca-1 and CD44 on wrist-level transverse sections from E14.5 ScxGFP -carrying wild-type embryos. Robust expression of ( B ) Sca-1 and ( D ) CD44 was detected in cells that surround the tendons at E14.5 (boxed areas), likely the precursors of the epitenon/paratenon. ( B’,D’ ) Higher magnification views of the boxed areas in ( B ) and ( D ). The epitenon/paratenon layer is indicated by white arrowheads. Note that both markers were not expressed by the tenocytes at E14.5, the onset of tenocyte differentiation or at any other stages during embryonic tendon development (not shown). Scale bars, 100 μm ( A–D ) and 25 μm ( B’,D’ ).

Article Snippet: Antibody , Goat anti-Sca-1/Ly6 , R and D Systems , Cat# AF1226 RRID: AB_354679 , IF(1:80).

Techniques: Immunofluorescence, Staining, Expressing

Journal: eLife

Article Title: Tgfβ signaling is critical for maintenance of the tendon cell fate

doi: 10.7554/eLife.52695

Figure Lengend Snippet:

Article Snippet: Antibody , Goat anti-Sca-1/Ly6 , R and D Systems , Cat# AF1226 RRID: AB_354679 , IF(1:80).

Techniques: Recombinant, In Situ, Staining

Journal: Cell stem cell

Article Title: Stress-induced changes in bone marrow stromal cell populations revealed through single-cell protein expression mapping

doi: 10.1016/j.stem.2019.06.003

Figure Lengend Snippet: Multi-Dimensional Single-Cell Mass Cytometry analysis reveals distinguishable clusters of bone marrow stromal cells

Article Snippet: Anti-Mouse Ly-6A/E (Sca-1) (D7)-164Dy , Fluidigm , Cat# 3164005B RRID:AB_2801436.

Techniques: Mass Cytometry, Purification, Affinity Purification, Functional Assay, Recombinant, Electron Microscopy, Modification, Conjugation Assay, Labeling, Isolation, RNA Library Preparation, Imaging, Gene Expression, Software