rabbit anti col1  (Bioss)

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

Journal: Frontiers in Immunology

Article Title: BTG2-deficient mast cells remodel the tumor and tumor-draining lymph node microenvironment leading to chemotherapy resistance in breast cancer

doi: 10.3389/fimmu.2025.1562700

Figure Lengend Snippet: Mast cells with low BTG2 expression induces the differentiation of αSMA-positive fibroblasts and enhances their collagen secretion through secreting tryptase to promote chemoresistance. (A) The percentage of different cancer-associated fibroblast subsets after chemotherapy between tumor and lymph node samples. (B) Representative images of αSMA immunofluorescent staining in BC tissues post-neoadjuvant chemotherapy between sensitive and resistant. Scale bar, 50 μm. (C) The correlation between the abundance of Tryptase + mast cells and αSMA + cells in tumor tissues of post-neoadjuvant chemotherapy (n = 94). Pearson’s correlation coefficient (R 2 ) and p values were determined by two-tailed Pearson correlation test. (D) Representative western blotting for phosphorylated/total ERK and aSMA in HFL-1 cells co-cultured with mast cells transfected with ctrl, shBTG2 or OE-BTG2. (E) Flow cytometry assay for the positive percentage of αSMA in HFL-1 cells co-cultured with mast cells transfected with ctrl, shBTG2 or OE-BTG2. (F) The positive percentage of aSMA in HFL-1 cells co-cultured with mast cells transfected with shBTG2 in culture medium with PBS, FSLLRY or GB88. (G) Differential gene expression analysis of fibroblast between sensitive or resistant to NAC group. The volcano plot displays log2 fold change (x-axis) versus -log10 p -value (y-axis) for all analyzed genes. Genes with a |log2 fold change| > 1 and a p -value < 0.05 are highlighted in red (upregulated) and blue (downregulated). The horizontal dashed line represents the significance threshold ( p < 0.05), while the vertical dashed lines indicate the fold change thresholds. (H) The correlation between the IHC score of COL1 and the abundance of Tryptase + mast cells in tumor tissues of post-neoadjuvant chemotherapy (n = 94). Pearson’s correlation coefficient (R 2 ) and p values were determined by two-tailed Pearson correlation test. (I) Representative western blotting for COL1 and COL3 in HFL-1 cells co-cultured with mast cells transfected with ctrl, shBTG2 or OE-BTG2. (J) Representative images of COL1 and Adriamycin immunofluorescent staining in BC tissues post-neoadjuvant chemotherapy between sensitive and resistant. Scale bar, 100 μm. ***p < 0.001 by Student's t-test.

Article Snippet: After blocking with TBS/0.05% Tween-20/5% BSA, the membranes were probed with first antibodies listed as below: rabbit anti-human p-FAK (Cat# 3283, Cell Signaling Technology; 1:1000), rabbit anti-human FAK (Cat# 3285, Cell Signaling Technology; 1:1000), rabbit anti-human p-ERK1/2 (Cat# ab201015, abcam; 1:1000), rabbit anti-human ERK1/2 (Cat# ab184699, abcam; 1:1000), rabbit anti-human Src (Cat# 2108S, Cell Signaling Technology; 1:1000), GAPDH (Cat# HRP-60004, Proteintech; 1:10,000), rabbit anti-human COL1 (Cat #72026, Cell Signaling Technology; 1:1000), rabbit anti-human COL3 (Cat #98908, Cell Signaling Technology; 1:1000), rabbit anti-human αSMA (Cat #19245, Cell Signaling Technology; 1:1000).

Techniques: Expressing, Staining, Two Tailed Test, Western Blot, Cell Culture, Transfection, Flow Cytometry, Gene Expression

Journal: iScience

Article Title: Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling

doi: 10.1016/j.isci.2025.112107

Figure Lengend Snippet: Rv1509 suppresses osteoblast differentiation and mineralization (A) ALP staining and subsequent quantification were conducted on days 7 and 14 while utilizing osteogenic differentiation medium. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗, p < 0.05; ∗∗, p < 0.01; [Student’s t test]). (B) Alizarin red staining and quantification were executed on days 14 and 21 in the context of osteogenic differentiation medium. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (C) RT-qPCR analysis was carried out to evaluate the expression levels of osteogenic genes in neonatal mouse calvarial cells that were treated with Rv1509 on days 1, 3, 7, and 14. The data are shown as mean ± SEM. ( n = 3; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (D) Representative western blots of Runx2, Osterix, and Col1 proteins in osteoblasts after Rv1509 treatment.

Article Snippet: Rabbit anti-Col1 Ab , ABclonal , Cat# A22089;RRID: AB_3665644.

Techniques: Staining, Quantitative RT-PCR, Expressing, Western Blot

Journal: iScience

Article Title: Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling

doi: 10.1016/j.isci.2025.112107

Figure Lengend Snippet: Rv1509 indirectly suppressed osteoblast differentiation by enhancing the release of inflammatory factors from osteoclasts (A) Alizarin red staining and quantitative assessment of osteoblasts cultured for a duration of 21 days with supernatants derived from both control and Rv1509-treated osteoclasts. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗, p < 0.05 [Student’s t test]). (B) Osteoblasts were subjected to treatment for 14 days with culture supernatants from control and Rv1509-treated osteoclasts, followed by ALP staining and subsequent quantification. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (C) The expression of osteoblast-specific mRNA was analyzed using RT-qPCR after a 7-day culture period with supernatants from control and Rv1509-treated osteoclasts. The data are shown as mean ± SEM. ( n = 3; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (D) ELISA was conducted to quantify the expression levels of inflammatory factors in osteoclasts from both the control and Rv1509-treated groups, while Sema4D expression was evaluated through RT-qPCR. The data are shown as mean ± SEM. ( n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (E) Representative protein blots illustrating the expression of Runx2, Osterix, and Col1 in osteoblasts.

Article Snippet: Rabbit anti-Col1 Ab , ABclonal , Cat# A22089;RRID: AB_3665644.

Techniques: Staining, Cell Culture, Derivative Assay, Control, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling

doi: 10.1016/j.isci.2025.112107

Figure Lengend Snippet: Rv1509 Induces osteoclast differentiation and RANKL secretion by osteoblasts via TLR2 (A) After inducing osteoclast differentiation for four days in the presence of Rv1509 and C29, TRAP staining (scale bar: 400 μm) and F-actin staining (scale bar: 100 μm) were performed, followed by quantitative analysis. The data are shown as mean ± SEM. ( n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (B) Following a four-day period of osteoclast differentiation stimulated by Rv1509 and C29, RT-qPCR was employed to evaluate the mRNA transcription levels of genes associated with osteoclasts. Additionally, ELISA was performed to measure the expression levels of inflammatory factors. The data are shown as mean ± SEM. ( n = 3; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (C) After 14 days of osteoblast differentiation induced by Rv1509 and C29, ALP staining was performed, followed by Alizarin red staining after 21 days. The data are shown as mean ± SEM. (scale bar: 1 mm; n = 3; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (D) The mRNA expression levels of osteoblast-related genes were analyzed using RT-qPCR after 14 days of differentiation in the presence of Rv1509 and C29. The data are shown as mean ± SEM. ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001 [Student’s t test]). (E) Protein blots showing the representative expression levels of Ctsk, NFATc1, MMP9, and c-Fos in osteoclasts treated with Rv1509 and C29. (F) Protein blots showing representative levels of Col1, Osterix, and Runx2 in osteoblasts after treatment with Rv1509 and C29.

Article Snippet: Rabbit anti-Col1 Ab , ABclonal , Cat# A22089;RRID: AB_3665644.

Techniques: Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing

Journal: iScience

Article Title: Mycobacterium tuberculosis specific protein Rv1509 modulates osteoblast and osteoclast differentiation via TLR2 signaling

doi: 10.1016/j.isci.2025.112107

Figure Lengend Snippet:

Article Snippet: Rabbit anti-Col1 Ab , ABclonal , Cat# A22089;RRID: AB_3665644.

Techniques: Virus, Recombinant, Lysis, Staining, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, BIA-KA, RNA Sequencing, Software

Journal: International journal of molecular sciences

Article Title: Focal Adhesion Kinase Alleviates Simulated Microgravity-Induced Inhibition of Osteoblast Differentiation by Activating Transcriptional Wnt/β-Catenin-BMP2-COL1 and Metabolic SIRT1-PGC-1α-CPT1A Pathways.

doi: 10.3390/ijms26041669

Figure Lengend Snippet: Figure 2. SMG attenuates OBD by inhibiting the Wnt/β-catenin signaling pathway. Western blot analysis of the steady-state levels of pFAK (Y397), COL1, β-catenin, ALP and BMP2 in MC3T3-E1 cells cultured under 1 g, SMG or SMG + CNF1 conditions. The histogram bars represent protein abundance normalized to the β-actin control and expressed relative to MC3T3-E1 cells cultured under the 1 g condition. Data from three independent experiments are presented as the mean ± SD. Significance analysis of the experimental data for each group was performed using Student’s t test (* p < 0.05; ** p < 0.01).

Article Snippet: Primary antibodies against SIRT1 (9475), β-catenin (8480), pFAK (pFAK-Y397) (3283), vinculin (13901) and COL1 (72026) were obtained from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Cell Culture, Quantitative Proteomics, Control