Journal: Advanced Science

Article Title: Discovery and Application of Postnatal Nucleus Pulposus Progenitors Essential for Intervertebral Disc Homeostasis and Degeneration

doi: 10.1002/advs.202104888

Figure Lengend Snippet: RegNPs were a metabolically active and mechanically sensitive population, while HomNPs were sensitive to hypoxia with “degenerative” potential. A) t‐SNE plots and representative violin plots showing the expression of Unc5c, Runx3, Bmp7, Wnt4, Tgfb2, CNTFR, Matn3, Grb10, Fgfr3, Epyc, Ptch1, and Pth1r on the t‐SNE map. B) Representation analysis of GO categories showing different functions for RegNPs. C) Heatmap revealing metabolic‐related functions and pathways for RegNPs. D) Representative images of lumbar spine sections from 4‐week‐old wild‐type mice stained for BMP7. Scale bars, 100 µm ( n = 3 mice per group). E) t‐SNE plots and representative violin plots showing the expression of Gdf5, Agt, Eln, Grem1, Mmp3, and Plau on the t‐SNE map. F) Representative analysis of GO categories showing different functions for HomNPs. G) Representative images of lumbar spine sections from 4‐week‐old WT mice stained for GDF5. The lower image shows a high‐magnification view of the indicated area from the upper image. Scale bars, 100 µm ( n = 3 mice per group).

Article Snippet: Dissociated single cells were then stained with AF488 anti‐mouse UTS2R (R&D Systems, FAB9245G‐100UG), APC anti‐mouse GDF5 (ASSAYPRO, 32579‐05161), anti‐mouse CNTFRa (Santa Cruz Biotechnology, SC9993), BB700 anti‐mouse CD146 (BD Biosciences, 742 280), APC anti‐mouse CD44 (BioLegend, 559 250), BV421 anti‐mouse CD73 (BioLegend, 127 217), BV605 anti‐mouse CD90 (BioLegend, 140 317), APC anti‐mouse PDGFa/CD140a (BioLegend, 135 907), FITC anti‐mouse Sca‐1 (BioLegend, 108 106), PerCP/Cy5.5 anti‐CD31 (BioLegend, 102 420), PerCP/Cy5.5 anti‐CD45 (BioLegend, 103 132), PerCP/Cy5.5 anti‐mouse TER‐119 (BioLegend, 116 228), FITC anti‐mouse 6C3/Ly‐51 (BioLegend, 108 305), Brilliant Violet 605 anti‐mouse CD90.2 (BioLegend, 140 317), PE/Cy7 anti‐mouse CD105 (BioLegend, 120 409), APC anti‐mouse CD200 (BioLegend, 123 809), and anti‐mouse Alexa Fluor 488 (Molecular Probes, A21202, 1:1000).

Techniques: Metabolic Labelling, Expressing, Staining

Journal: Advanced Science

Article Title: Discovery and Application of Postnatal Nucleus Pulposus Progenitors Essential for Intervertebral Disc Homeostasis and Degeneration

doi: 10.1002/advs.202104888

Figure Lengend Snippet: Lineage tracing of UTS2R + NP cells. A) Dot plot showing the expression of Uts2r on the t‐SNE map. B) Representative immunofluorescence imaging of UTS2R (green) in postnatal 1‐month‐old WT mice. The right image shows high magnification of the indicated area from the left image ( n = 3 mice per group). Scale bars, 100 µm. C) Construction strategy of Uts2r‐CreER transgenic mice using the CRISPR/Cas9 System. D) Diagram showing postnatal day 1 (P1) Uts2r‐CreER;Ai9 /+ mice administered with one dosage tamoxifen and sacrificed at postnatal day 3 (P3), 1 month (P1M), or 2 months (P2M). E) Representative immunofluorescence imaging of Uts2r‐CreER;Ai9 + cells (red). The right images show high magnification of the indicated area from the left image ( n = 3 mice per group). Scale bars, 100 µm. F,G) Representative immunofluorescence imaging of Uts2r‐CreER;Ai9 + cells (red), BMP7 (green) (F) or GDF5 (green) (G). The right images show high magnification of the indicated area from the left image ( n = 3 mice per group). Scale bars, 100 µm.

Article Snippet: Dissociated single cells were then stained with AF488 anti‐mouse UTS2R (R&D Systems, FAB9245G‐100UG), APC anti‐mouse GDF5 (ASSAYPRO, 32579‐05161), anti‐mouse CNTFRa (Santa Cruz Biotechnology, SC9993), BB700 anti‐mouse CD146 (BD Biosciences, 742 280), APC anti‐mouse CD44 (BioLegend, 559 250), BV421 anti‐mouse CD73 (BioLegend, 127 217), BV605 anti‐mouse CD90 (BioLegend, 140 317), APC anti‐mouse PDGFa/CD140a (BioLegend, 135 907), FITC anti‐mouse Sca‐1 (BioLegend, 108 106), PerCP/Cy5.5 anti‐CD31 (BioLegend, 102 420), PerCP/Cy5.5 anti‐CD45 (BioLegend, 103 132), PerCP/Cy5.5 anti‐mouse TER‐119 (BioLegend, 116 228), FITC anti‐mouse 6C3/Ly‐51 (BioLegend, 108 305), Brilliant Violet 605 anti‐mouse CD90.2 (BioLegend, 140 317), PE/Cy7 anti‐mouse CD105 (BioLegend, 120 409), APC anti‐mouse CD200 (BioLegend, 123 809), and anti‐mouse Alexa Fluor 488 (Molecular Probes, A21202, 1:1000).

Techniques: Expressing, Immunofluorescence, Imaging, Transgenic Assay, CRISPR

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Relative mRNA expressions of Bmp15, Gdf9, and C-KIT in the ovaries of mice aged from 10 to 40 weeks. Levels of the mRNAs of Bmp15, Gdf9, and C-KIT were normalized to the amount of GAPDH per sample. Data are presented as mean ± standard deviation.*p < 0.05 (vs. 10 weeks old).

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Immunohistochemistry of BMP15, GDF9, and C-KIT in the mouse ovaries according to age. (a, e, i) 10 weeks, (b, f, j) 20 weeks, (c, g, k) 30 weeks, and (d, h, l) 40 weeks of mice. Immunostainings with anti-BMP15 (a–d) and cv-Kit (i–l) antibodies were usually localized in oocyte and cumulus cells, whereas immunostaining with anti-GDF antibody (e–h) was detected just in the oocytes (brown). (m) Quantitative results of immunohistochemistry analysis. Five ovaries were used per age group and a total of 10 follicles per ovary were counted for analysis. The images are representative at a final magnification of ×100. Scale bar indicates 50 µm. Black arrowhead, white arrowhead, black arrow, and red arrow indicate oocytes of primary, secondary, antral, and graffian follicles, respectively. *p < 0.05 (vs. 10 weeks old). (A color version of this figure is available in the online journal.)

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Immunohistochemistry, Immunostaining

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Protein expressions of BMP15, GDF9, and C-KIT in the ovaries of mice aged 10 to 40 weeks. Protein expression levels of BMP15, GDF9, and C-KIT were normalized to the amount of actin per sample. Relative protein densities were quantified using NIH-Image J software (version 1.35d). Data are presented as mean ± standard deviation. Results are representative of at least three independent experiments. *p < 0.05 (vs. 10 weeks old).

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: Expressing, Software, Standard Deviation

Journal: Experimental Biology and Medicine

Article Title: Prediction of ovarian aging using ovarian expression of BMP15, GDF9, and C-KIT

doi: 10.1177/1535370220915826

Figure Lengend Snippet: Expressions changes of Bmp15, Gdf9 and C-KIT in transcriptome analysis using RNA-Seq for ovaries of mice aged 6 weeks and 48 weeks. (a) Heat-map results, (b) quantification of expression level of mRNA. Expressions of BMP15, GDF9, and C-KIT were significantly decreased in old mice compared with young mice.

Article Snippet: The sections were incubated with antiserum at a dilution of 1:200 in BMP15 (ORB247897, Biorbyt, Cambridge, UK), 1: 100 in GDF9 (AF739, R&D Systems, MN, USA), and 1:100 in C-KIT ( {"type":"entrez-protein","attrs":{"text":"ORB10286","term_id":"1177131250","term_text":"ORB10286"}} ORB10286 , Biorbyt) for overnight.

Techniques: RNA Sequencing, Expressing