mouse anti col1  (Proteintech)

Journal: Cell reports

Article Title: PAI-1 uncouples integrin-β1 from restrain by membrane-bound β-catenin to promote collagen fibril remodeling in obesity-related neoplasms

doi: 10.1016/j.celrep.2024.114527

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-COL1 (E3E1X) , Cell Signaling Technology , Cat# 66948S; RRID: AB_2920541.

Techniques: Recombinant, Membrane, Cell Culture, Reverse Transcription, SYBR Green Assay, Clinical Proteomics, Protein Extraction, Extraction, Bicinchoninic Acid Protein Assay, In Situ, Blocking Assay, Migration, shRNA, Control, Software, Pyromark Assay, Western Blot, Simple Western

Journal: Cell reports

Article Title: PAI-1 uncouples integrin-β1 from restrain by membrane-bound β-catenin to promote collagen fibril remodeling in obesity-related neoplasms

doi: 10.1016/j.celrep.2024.114527

Figure Lengend Snippet:

Article Snippet: For DKK1-, SPARC-, TIMP2-, COL1, integrin β2, or integrin α3 PLA, purified rabbit monoclonal anti-DKK1 (1:200, Cell Signaling Technology, Cat# 48367S), rabbit polyclonal anti-SPARC (1:200, Cell Signaling Technology, Cat# 5420S), mouse monoclonal anti-TIMP2 (1:200, R&D Systems, Cat# MAB971), mouse monoclonal anti-COL1 (1:200, Cell Signaling Technology, Cat# 66948S), goat polyclonal anti-integrin β1 (1:200, R&D Systems, Cat# AF1778), goat polyclonal anti-integrin β2 (1:200, R&D Systems, Cat# AF1730), and mouse monoclonal anti-integrin α3 (1:200, R&D Systems, Cat# MAB1345) were used.

Techniques: Recombinant, Membrane, Cell Culture, Reverse Transcription, SYBR Green Assay, Clinical Proteomics, Protein Extraction, Extraction, Bicinchoninic Acid Protein Assay, In Situ, Blocking Assay, Migration, shRNA, Control, Software, Pyromark Assay, Western Blot, Simple Western

Journal: Advanced Science

Article Title: WAC Facilitates Mitophagy‐mediated MSC Osteogenesis and New Bone Formation via Protecting PINK1 from Ubiquitination‐Dependent Degradation

doi: 10.1002/advs.202404107

Figure Lengend Snippet: WAC was up‐regulated during the osteogenic differentiation of MSCs. a) ARS Staining and quantification throughout MSC osteogenic differentiation; b) ALP Staining and quantification during the progression of MSC osteogenesis; c) Immunofluorescence Staining for COL1 (red) during osteogenic induction. Quantification is presented in the right panel (Scale bar = 50 µm); d) Relative mRNA levels of WAC assessed by qRT‐PCR at different time points during MSC osteogenesis; e) Protein levels of WAC and osteogenic markers (RUNX2, Osterix, OCN) during MSC osteogenic differentiation; f) Pearson correlation analysis depicting the relationship between WAC expression and quantification of RUNX2, Osterix, and OCN levels during MSC osteogenic differentiation; g) qRT–PCR and Western blotting to detect WAC mRNA and protein levels in bone marrow MSCs from nonosteoporotic patients and patients with osteoporosis; h) HE staining and immunohistochemical staining for WAC in the femurs of SAMR1 mice and SAMP6 mice (Scale bar = 100 µm). All data are presented as the means ± SD, n = 6 per group in (a, b, c), n = 5 in (g), n = 9 in (d, e, g), n = 12 in (f). Statistical differences were determined using Student's t ‐test or ANOVA. ** p < 0.01 and *** p < 0.001.

Article Snippet: Subsequently, the cells were treated with Triton X‐100 for 15 min at room temperature and incubated with goat serum for 30 min. Primary antibodies against COL1 (Cell Signaling Technology, Cat. No. 66 948), LC3B (Abcam, Cat. No. Ab192890), or TOM20 (Abcam, Cat. No. Ab186735) were added and left to incubate overnight at 4 °C.

Techniques: Staining, Immunofluorescence, Quantitative RT-PCR, Expressing, Western Blot, Immunohistochemical staining

Journal: Advanced Science

Article Title: WAC Facilitates Mitophagy‐mediated MSC Osteogenesis and New Bone Formation via Protecting PINK1 from Ubiquitination‐Dependent Degradation

doi: 10.1002/advs.202404107

Figure Lengend Snippet: WAC positively regulated the osteogenic differentiation of MSCs in vitro. WAC is modulated in MSCs through SiRNA and Lentivirus. a) MSCs were cultured in osteogenic medium after transfection with SiRNA. ARS staining, ALP staining, and quantification were performed on day 12; b) Western blotting for protein levels of osteogenesis‐related markers (RUNX2, Osterix, OCN). Quantification is presented in the right panel; c) Immunofluorescence staining for COL1 (red) after SiRNA transfection. Quantification is shown in the right panel (Scale bar = 50 µm); d) Overexpression lentivirus of WAC transfected into MSCs, followed by culture in osteogenic medium. ARS staining, ALP staining, and quantification conducted on day 12; e) Western blotting for protein levels of osteogenesis‐related markers after overexpression lentivirus transfection; f) Immunofluorescence staining for COL1 (red) after overexpression lentivirus transfection. Quantification is shown in the right panel (Scale bar = 50 µm). All data are presented as the means ± SD, n = 6 per group in (a, c, d, f), n = 9 per group in (b, e). Statistical differences were determined using Student's t ‐test or ANOVA. ns not statistically significant, ** p < 0.01 and *** p < 0.001.

Article Snippet: Subsequently, the cells were treated with Triton X‐100 for 15 min at room temperature and incubated with goat serum for 30 min. Primary antibodies against COL1 (Cell Signaling Technology, Cat. No. 66 948), LC3B (Abcam, Cat. No. Ab192890), or TOM20 (Abcam, Cat. No. Ab186735) were added and left to incubate overnight at 4 °C.

Techniques: In Vitro, Cell Culture, Transfection, Staining, Western Blot, Immunofluorescence, Over Expression

Journal: Tissue Engineering and Regenerative Medicine

Article Title: In Vitro Assessment of Injectable Bone Marrow Aspirate Concentrates Compared to Injectable Platelet-Rich Fibrin

doi: 10.1007/s13770-024-00677-7

Figure Lengend Snippet: List of primer sequences for real-time PCR

Article Snippet: The RGFs were fixed with 4% formaldehyde for 10 min, permeabilized with PBS containing 0.2% Triton X-100, and blocked with PBS containing 1% bovine serum albumin (BSA; Sigma Aldrich) for 1 h. Subsequently, the cells were incubated overnight at 4 °C with an anti-mouse monoclonal COL1 antibody (MA1-26771; Invitrogen, Waltham, MA, USA) diluted 1:200 in PBS containing 1% BSA.

Techniques: Sequencing

Journal: Tissue Engineering and Regenerative Medicine

Article Title: In Vitro Assessment of Injectable Bone Marrow Aspirate Concentrates Compared to Injectable Platelet-Rich Fibrin

doi: 10.1007/s13770-024-00677-7

Figure Lengend Snippet: A Real-time PCR was performed to determine the relative mRNA levels of transforming growth factor-beta (TGF-β), vascular endothelial growth factor (VEGF) and collagen I (COL1) in RGFs cultured with control, iPRF, or iBMAC at 3 and 7 days postseeding. B Representative immunofluorescence staining of COL1 (green) with DAPI staining (blue) of RGFs at 14 days and C the quantified values of COL1 staining in comparison with each group. (# denotes significantly lower than all other modalities, p < 0.05; and ** denotes values significantly greater than those of all other treatment modalities, p < 0.05)

Article Snippet: The RGFs were fixed with 4% formaldehyde for 10 min, permeabilized with PBS containing 0.2% Triton X-100, and blocked with PBS containing 1% bovine serum albumin (BSA; Sigma Aldrich) for 1 h. Subsequently, the cells were incubated overnight at 4 °C with an anti-mouse monoclonal COL1 antibody (MA1-26771; Invitrogen, Waltham, MA, USA) diluted 1:200 in PBS containing 1% BSA.

Techniques: Real-time Polymerase Chain Reaction, Cell Culture, Control, Immunofluorescence, Staining, Comparison

Journal: Tissue Engineering and Regenerative Medicine

Article Title: In Vitro Assessment of Injectable Bone Marrow Aspirate Concentrates Compared to Injectable Platelet-Rich Fibrin

doi: 10.1007/s13770-024-00677-7

Figure Lengend Snippet: A Real-time PCR was performed to determine the relative mRNA levels of runt-related transcription factor 2 (Runx2), COL1 and osteocalcin (OCN) after 3 and 14 days of culture with control, iPRF or iBMAC in ROBs. B Representative alizarin red staining of ROBs 14 days after cell seeding and C the quantified values. (* denotes a statistically significant difference between groups, p < 0.05; # denotes a significantly lower value than that of all the other modalities, p < 0.05; and ** denotes a significantly greater value than that of all other treatment modalities, p < 0.05)

Article Snippet: The RGFs were fixed with 4% formaldehyde for 10 min, permeabilized with PBS containing 0.2% Triton X-100, and blocked with PBS containing 1% bovine serum albumin (BSA; Sigma Aldrich) for 1 h. Subsequently, the cells were incubated overnight at 4 °C with an anti-mouse monoclonal COL1 antibody (MA1-26771; Invitrogen, Waltham, MA, USA) diluted 1:200 in PBS containing 1% BSA.

Techniques: Real-time Polymerase Chain Reaction, Control, Staining