mouse col1 mab  (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: Purinergic Signalling

Article Title: Modafinil exerts anti-inflammatory and anti-fibrotic effects by upregulating adenosine A 2A and A 2B receptors

doi: 10.1007/s11302-023-09973-8

Figure Lengend Snippet: Expression amounts of ARs and ACs are altered in fibrotic diseases Protein amounts of ARs (a-e) and Col (f) were measured by western blotting. a Expression amounts of A2AAR and A2BAR in NHLFs, DHLFs, and TGFβ (2ng/ml)-treated NHLFs. The blots shown are representative of three to five experiments performed using three to five different cultures. b Expression amounts of A2AAR in liver and lung tissues from wild-type and murine models of TAA-induced liver fibrosis or BLM-induced lung fibrosis. c Expression amounts of A2BAR in liver and lung tissues from wild-type and murine models of TAA-induced liver fibrosis or BLM-induced lung fibrosis. d and e Expression levels of A1AR (d) and A3AR (e) in the liver and lung tissues from wild-type and murine models of TAA-induced liver fibrosis or BLM-induced lung fibrosis, respectively. f Change in Col1 content of NHLFs transfected with scrambled siRNA or A2R-siRs. g Messenger RNA levels of ACs in NHLFs and TGFβ-treated NHLFs. NHLFs were treated with TGFβ for 24 h, and mRNA levels of ACs were measured by RT-PCR. Three to seven tissues per mice group were employed. Each experiment was repeated three times independently. Data are presented as the mean ± SEM values. *P < 0.05, **P < 0.01 vs. NHLFs or control

Article Snippet: The membranes were immunoblotted using antibodies against A 1 AR (ab151523, Abcam), A 2A AR (ab3461, Abcam), A 2B AR (ab229671, Abcam) and A 3 AR (ab197350, Abcam), K Ca 3.1 (sc-365,265, Santa Cruz Biotechnology, Dallas, TX, USA), K Ca 2.3 (sc28621, Santa Cruz Biotechnology), Col1 (#66,948, Cell Signaling Technology Inc., Danvers, MA, USA), α-SMA (ab7817, Abcam), Epac1 (sc-28,366, Santa Cruz Biotechnology), and Epac2 (sc28326f, Santa Cruz Biotechnology), and bands were visualized by chemiluminescence.

Techniques: Expressing, Western Blot, Transfection, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Purinergic Signalling

Article Title: Modafinil exerts anti-inflammatory and anti-fibrotic effects by upregulating adenosine A 2A and A 2B receptors

doi: 10.1007/s11302-023-09973-8

Figure Lengend Snippet: cAMP signaling is reduced in fibrotic diseases a Expression amounts of Epac1 in liver tissues from wild-type and murine models of liver fibrosis. Epac1 was significantly downregulated in liver fibrosis models compared to the control. Three to eight tissues per mice group were employed. Each experiment was repeated three times independently. b Expression amounts of Epac2 in NHLFs. Treatment with TGFβ (10 ng/ml) for 24 h reduced Epac2 levels. c Expression amounts of Epac1, Epac2, α-SMA, and Col1 in NHLFs and DHLFs. Epac1 and Epac2 amounts were significantly reduced, and α-SMA and Col1 amounts were significantly elevated in DHLFs compared to the NHLFs. d and e Expression amounts of Col1 (d) and α-SMA (e) in NHLFs. TGFβ (10 ng/ml) augmented expression amounts of Col1 and α-SMA. Augmented amounts of Col1 and α-SMA were reversed using cAMP-elevating agents: NECA (100 nM) or MF-S. b-e Blots shown are representative of the three to five experiments performed using three to five different cultures. Data are presented as mean ± SEM values. *P < 0.05, **P < 0.01 vs. control. #P < 0.05, ##P < 0.01 vs. DHLFs or TGFβ-treated NHLFs.

Article Snippet: The membranes were immunoblotted using antibodies against A 1 AR (ab151523, Abcam), A 2A AR (ab3461, Abcam), A 2B AR (ab229671, Abcam) and A 3 AR (ab197350, Abcam), K Ca 3.1 (sc-365,265, Santa Cruz Biotechnology, Dallas, TX, USA), K Ca 2.3 (sc28621, Santa Cruz Biotechnology), Col1 (#66,948, Cell Signaling Technology Inc., Danvers, MA, USA), α-SMA (ab7817, Abcam), Epac1 (sc-28,366, Santa Cruz Biotechnology), and Epac2 (sc28326f, Santa Cruz Biotechnology), and bands were visualized by chemiluminescence.

Techniques: Expressing, Control

Journal: Purinergic Signalling

Article Title: Modafinil exerts anti-inflammatory and anti-fibrotic effects by upregulating adenosine A 2A and A 2B receptors

doi: 10.1007/s11302-023-09973-8

Figure Lengend Snippet: MF-S inhibits fibrotic responses in murine models of fibrotic diseases a and b Hydroxyproline or P1NP levels in murine models of TAA-induced liver fibrosis or BLM-induced lung fibrosis. a Hydroxyproline levels in liver tissue from mice treated with TAA for 8 or 16 weeks. b Hydroxyproline or P1NP levels were elevated in models of fibrotic diseases, and elevated levels of hydroxyproline or P1NP were significantly reversed after MF-S treatment. c and d Expression amounts of Col1 (c) or α-SMA (d) in TAA-induced liver fibrosis and BLM-induced lung fibrosis murine models. e Representative images of Masson’s trichrome, Sirus red, reticulin, and α-SMA staining in lung tissues from mice with BLM-induced pulmonary fibrosis. Enlargements of black boxes (solid line) were shown in right panels. Three to ten tissues per mice group were employed. Each experiment was repeated three to five times independently. Data are presented as mean ± SEM values. *P < 0.05, **P < 0.01 vs. control. #P < 0.05, ##P < 0.01 vs. BLM-induced lung disease model

Article Snippet: The membranes were immunoblotted using antibodies against A 1 AR (ab151523, Abcam), A 2A AR (ab3461, Abcam), A 2B AR (ab229671, Abcam) and A 3 AR (ab197350, Abcam), K Ca 3.1 (sc-365,265, Santa Cruz Biotechnology, Dallas, TX, USA), K Ca 2.3 (sc28621, Santa Cruz Biotechnology), Col1 (#66,948, Cell Signaling Technology Inc., Danvers, MA, USA), α-SMA (ab7817, Abcam), Epac1 (sc-28,366, Santa Cruz Biotechnology), and Epac2 (sc28326f, Santa Cruz Biotechnology), and bands were visualized by chemiluminescence.

Techniques: Expressing, Staining, Control