type 1 collagen col1  (Thermo Fisher)

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Biological functional analysis of tri-layer scaffold systems. A) Representative images of migrated cells stained with crystal violet at 48 h. B) Number of migrating cells in control and CE-SKP. C) Glycosaminoglycan concentrations in control and CE-SKP at 7 and 14 days. D) qPCR analysis for COL2, SOX9, and ACAN. E) Western blotting of COL2 and SOX9. F) Alizarin red staining of control and CPH. G) ALP activity of control and CPH. H) qPCR analysis for COL1, COL10, OCN, and RUNX2. I) Western blotting of COL10, COL1, and RUNX2. J) Representative images of migrated cells stained with crystal violet at 48 h. K) Number of migrating cells in control and P2G3. L) Scanning electron microscope micrographs of P2G3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Functional Assay, Staining, Control, Western Blot, Activity Assay, Microscopy

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Regeneration of the femoral trochlea in a rabbit model. Representative images of IHC staining for A) COL1, B) COL10.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Immunohistochemistry

Journal: Bone & Joint Research

Article Title: High-intensity running exercise promotes knee meniscal damage via the PI3K/AKT/mTOR axis

doi: 10.1302/2046-3758.1411.BJR-2024-0535.R1

Figure Lengend Snippet: Excessive mechanical stress promotes meniscus degeneration.a) Immunohistochemical staining revealed the expression of Aggrecan and matrix metalloproteinase 13 (MMP13) in the anterior horn, posterior horn, and articular surface of the meniscus following treadmill intervention in mice. Scale bar: 100 µm. (The same group showed serial sections from the same mouse knee joint tissue for staining analysis.) b) Statistical analysis of the differences in MMP13 and Aggrecan expression between the anterior and posterior horns of the meniscus and the articular cartilage surface groups (n = 5/group). Data were analyzed using one-way analysis of variance (ANOVA) followed by Tukey's post-hoc test. c) Immunofluorescence displaying COL1-positive, Aggrecan-positive, MMP13-positive, and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5)-positive cells as a proportion of total meniscal cells in primary medial meniscal cells (n = 3/group). Data are presented as the mean (SD). Scale bar: 200 µm. d) Quantification of the positive meniscus cells (P-PI3K, P-AKT, and P-S6) as a proportion of total meniscal cells (n = 3/group). Data were analyzed using independent-samples t -test. *p < 0.05, **p < 0.01, ***p < 0.001. ns, non-significant.

Article Snippet: Mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:10,000, ab8245; Abcam, UK), rabbit anti-MMP13 (1:2,000, 18165-1-AP; Proteintech, USA), rabbit anti-A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) (1:1,000, A2836; Abclonal, USA), rabbit anti-collagen type I (COL1) (1:2,000, 14695-1-AP; Proteintech), rabbit anti-Aggrecan (1:1,000, A11691; Abclonal), rabbit anti-phospho-PI3K p85 (1:1,000, YP0224; ImmunoWay, USA), rabbit anti-phospho-AKT (Thr308) (1:500, PC2720; Abmart, China), rabbit anti-phospho-S6 (S240/S244) (1:500, AP0537; Abclonal), rabbit anti-PI3K (1:500, A17433; Abclonal), rabbit anti-AKT (1:1,000, ab192623; Abcam), and rabbit anti-S6 (1:500, A6058; Abclonal) served as the primary antibodies used.

Techniques: Immunohistochemical staining, Staining, Expressing, Immunofluorescence