anti col1 primary antibody  (Proteintech)

Journal: Communications Biology

Article Title: GDF11 alleviates glucocorticoid-induced osteonecrosis of the femoral head by regulating angiogenesis via the PI3K-AKT-eNOS pathway

doi: 10.1038/s42003-025-09078-5

Figure Lengend Snippet: A , B Angiography of femoral heads and subsequent vessel volume quantification in the experimental groups: sham, MP, MP + AAV-NC, MP + AAV-GDF11 (n = 6 per group). C Representative transverse sections of H&E and Masson staining from the sham, MP, MP + AAV-NC, and MP + AAV-GDF11 groups are shown. Scale bar = 500 μm. D Quantification of angiography particles number in different groups (n = 6 per group). E Representative immunofluorescence images of the sham, MP, MP + AAV-NC, and MP + AAV-GDF11 groups showing CD31 (green), COL1 (red), and DAPI-stained nuclei (blue). Scale bar = 100 μm. F , G Quantitative analysis of mean fluorescence in different groups (n = 6 per group). H , I IHC staining for CD31 and quantitative analysis of CD31-positive cells in the sham, MP, MP + AAV-NC, and MP + AAV-GDF11 groups (n = 6 per group). Scale bar = 100 μm (upper panel) and 50 μm (lower panel). Significant differences are indicated as follows: ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: Primary antibodies like COL1 (#72026, Cell Signaling Technology), VEGFA (ab52917, Abcam), CD31 (ab182981, Abcam), p-PI3K (TA3242, Abmart), p-eNOS (PC3447, Abmart) and corresponding fluorescent secondary antibodies (ab150079 and ab150077, Abcam) were used.

Techniques: Staining, Immunofluorescence, Fluorescence, Immunohistochemistry

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: Bioactive Materials

Article Title: Sustained release of magnesium and zinc ions synergistically accelerates wound healing

doi: 10.1016/j.bioactmat.2023.02.019

Figure Lengend Snippet: Mechanism via which GelMA/Mg/Zn promotes soft tissue wound healing. (A) Volcano map showing differentially expressed genes in the control (Ctrl) and GelMA/Mg/Zn groups. (B) Hierarchical gene clustering of RNA-Seq results showing that various genes related to biological processes were different between the Ctrl and GelMA/Mg/Zn groups. (C) Representative top 20 upregulated or downregulated KEGG pathways. In the bubble diagram, the x-axis represents the enrichment score of the gene sets, the y-axis shows gene set names, and the size of the bubble indicates the number of genes in that gene set. (D) A representative Western blot shows the levels of α-SMA, COL1, STAT3, and p-STAT3 in human skin fibroblasts from the Ctrl, GelMA, GelMA/Mg, GelMA/Zn, and GelMA/Mg/Zn groups, treated and untreated with the Stattic STAT3 inhibitor.

Article Snippet: The membrane was incubated overnight with specific primary antibodies COL1 (Proteintech, Chicago, IL, USA), α-SMA (Proteintech), STAT3 (Huaxingbochuang, Beijing, China), P-STAT3 (Huaxingbochuang), or GAPDH (Cell Signaling Technology, Beverly, MA, USA) at 4 °C after sealing with 10% nonfat milk.

Techniques: Control, RNA Sequencing, Western Blot

Journal: PLoS ONE

Article Title: Effects of Negative Pressure Wound Therapy on Mesenchymal Stem Cells Proliferation and Osteogenic Differentiation in a Fibrin Matrix

doi: 10.1371/journal.pone.0107339

Figure Lengend Snippet: Immediately before and 3, 7 days after incubation, cell clots were harvested and confocal microscopy was performed to detect fluorescent immunostaining for COL1. Results showed that the expression of COL1 was absent or weak in two groups before incubation, but stronger in NPWT group at day 3 and 7 after incubation. Bars = 50 µm.

Article Snippet: Then they were blocked with goat serum, and incubated with the specific rat primary antibodies against collagen type 1 (COL1) (1∶100, BOSTER, Wuhan, China) overnight at 4°C.

Techniques: Incubation, Confocal Microscopy, Immunostaining, Expressing

Journal: PLoS ONE

Article Title: Effects of Negative Pressure Wound Therapy on Mesenchymal Stem Cells Proliferation and Osteogenic Differentiation in a Fibrin Matrix

doi: 10.1371/journal.pone.0107339

Figure Lengend Snippet: NPWT-treated and static control group cell clots were harvested before and 1, 3, and 7 days after incubation (n = 3 per timepoint for each group) and analyzed by real-time RT-PCR. The expressions of the specific osteogenic genes (A) ALP, (B) OC, and (D) COL1 were higher in NPWT group than those in control group (* p <0.05, ** p <0.01). The NPWT group also expressed higher levels of the osteogenic transcription factor Cbfa1 (also known as Runx2) and the mechanosignaling molecule integrin β5 (ITGB5) (C) and (E) (* p <0.05, ** p <0.01).

Article Snippet: Then they were blocked with goat serum, and incubated with the specific rat primary antibodies against collagen type 1 (COL1) (1∶100, BOSTER, Wuhan, China) overnight at 4°C.

Techniques: Control, Incubation, Quantitative RT-PCR