anti collagen type x  (Thermo Fisher)

Journal: Advanced Healthcare Materials

Article Title: Start, Stop, Rewind, Repeat—Cyclic Exposure of Adipose Stromal Cells‐derived Cartilage Organoids to Chondrogenic and Proliferative Cues to Achieve Scaled‐up and Customizable Bone Formation by Endochondral Ossification

doi: 10.1002/adhm.202504880

Figure Lengend Snippet: Implanted human‐ and host recruited murine‐cells contribution to endochondral ossification and bone remodeling. ((Human implanted cells initiate bone formation both on the outer and core parts of the Pa‐TEC while mouse recruited cells create bone marrow compartments and mediate bone remodeling over time. Immuno‐histochemistry staining of collagen type X (Col10), human collagen type I (hCol1) and human Nuclei (huNu) in Pa‐TEC (adult donors) after (A) 4 weeks, (B) 12 weeks and (C) 24 weeks of implantation. (D) Human and mouse nuclei quantification in outer and core areas of the implanted Pa‐TECs performed on huNu stained images. Data are expressed in percentage of nuclei (mean ± SD). (N ≥ 15 images per time point per area, 1–2 biological replicate per donor, 3 adult donors tested). (E) Evolution and quantification of hCol1 in bone tissues of the implanted Pa‐TECs after 12 and 24 weeks performed on hCol I stained images. Data are expressed in percentage of bone tissue (mean ± SD). (N ≥ 5 Pa‐TEC per time point, 1–2 biological replicate per donor, 5 adult donors tested). For statistical analyses one‐way ANOVA with Tukey's multiple comparisons tests were used. Blue arrow indicates mouse cells (blue nuclei), black arrows indicates human cells (brown nuclei). Red scale bar = 100 µm. Symbols, cartilage (c), bone (b), immature bone (b*), bone marrow (BM) and human (h). Yellow dotted lines indicate bone or immature bone areas.)).

Article Snippet: The following primary antibodies were diluted at 1:200 (unless otherwise specified) in blocking buffer: anti‐human Collagen Type X (Invitrogen, #14‐9771‐82, dilution 1:100), anti‐human MMP13 (Abcam, #ab39012), anti‐human Cleaved Caspase‐3 (Abcam, #ab39012), anti‐human Collagen Type II (Merck Millipore, #MAB8887), anti‐human Laminin (Abcam, #ab11575) and anti‐human Fibronectin (Abcam, #ab2413).

Techniques: Immunohistochemistry, Staining

Journal: Bone & Joint Research

Article Title: Genotype and phenotype correlation analysis in 27 families with multiple osteochondroma and validation by ATDC5 chondrocyte models

doi: 10.1302/2046-3758.1410.BJR-2024-0477.R1

Figure Lengend Snippet: The evaluation of chondrocyte differentiation. The expression of differentiation markers of chondrocytes (α-1 chain of type X collagen ( COL10A1 ), matrix metallopeptidase 13 ( MMP13 ), and Runt-related transcription factor 2 ( RUNX2 )), as measured by a) to c) quantitative real-time polymerase chain reaction (qRT-PCR) and d) western blot, was significantly higher in the EXT1 -/- and EXT2 -/- groups compared with the wild type (WT) group. e) and f) The results of Alizarin red staining (magnification: 10×) on day 21 also showed that calcium deposition (stained red) was significantly increased in the EXT1 -/- and EXT2 -/- groups compared with the WT group. *p < 0.05, **p < 0.01; independent-samples t -test. Scale bar = 200 μm. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; KD, knockdown; mRNA, messenger RNA; ns, no significance.

Article Snippet: Membranes were probed with primary antibodies as follows: EXT1 (1:1,000 dilution, Thermo Fisher Scientific), EXT2 (1:1,000 dilution, Proteintech, USA), Aggrecan Monoclonal Antibody (BC-3) (ACAN) (1:1,000 dilution, Thermo Fisher Scientific), α-1 chain of type I collagen (COL1A1) (1:1,000 dilution, Cell Signaling Technology (CST), USA), SOX9 (1:5,000 dilution, Proteintech), RUNX2 (1:2,000 dilution, Proteintech), α-1 chain of type X collagen (COL10A1) (1:1,000 dilution, Proteintech), MMP13 (1:1,000 dilution, Proteintech), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (1:10,000 dilution, Proteintech), and Tubulin (1:5,000 dilution, Proteintech).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot, Staining, Knockdown