smad3 peptides  (GenScript corporation)

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: miR-552-3p regulates TGF-β1/Smad3 signaling pathway in LX-2 cells. (A) Relative mRNA levels of fibrotic genes in miR-552-3p (1 nM) or NC (1 nM)-transfected LX-2 cells treated with 10 ng/mL TGF-β1 or not for 24 h. (B) Relative mRNA levels of inflammatory genes in miR-552-3p (1 nM) or NC (1 nM)-transfected LX-2 cells treated with 800 ng/mL LPS or not for 8 h. (C-D) The protein levels of Collagen Ⅰ, p-Smad3, t-Smad3 and α-SMA in LX-2 cells transfected with miR-552-3p (1 nM) for 48 h and then treated with 10 ng/mL TGF-β1 or not for 24 h. (E) Relative mRNA levels of SMAD3 and TGFBR2 after the LX-2 cells were transfected with different concentrations of miR-552-3p (30, 10, 1, 0.1 nM) for 48 h. (F) MiR-552-3p level in LX-2 cells transfected with Anti-miR-552-3p or Anti-NC (50 nM) for 72 h. (G) Relative mRNA levels of SMAD3 and TGFBR2 after the LX-2 cells were transfected with Anti-miR-552-3p for 72 h. Data are presented as the mean ± SD of three independent experiments. *P<0.05, ** P<0.01, ***P<0.001 vs. NC or Anti-NC; #P<0.05, ##P <0.01, ###P <0.001 vs. H 2 O.

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Transfection

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: miR-552-3p down-regulates fibrotic and inflammatory responses in HFHFrHC diet-induced NASH mouse model. (A) Schematic diagram of HFHFrHC-induced animal experiment. (B) Relative level of miR-552-3p in the liver tissues. (C) HE staining of liver samples. Scale bar: 100 µm. (D) Liver coefficient presented by liver weight to body weight ratio. (E-F) Serum ALT and AST levels. (G) Sirius red, Masson's trichrome and α-SMA staining of liver samples. Scale bar: 100 µm. (H-J) Fibrotic area per field was quantified. (K) Content of hydroxyproline in mouse livers. (L-M) Relative mRNA levels of fibrotic and inflammatory genes in mouse livers. (N-O) Protein levels of Collagen Ⅰ, p-Smad3 and t-Smad3 in TGF-β1/Smad3 signaling pathway. (P) Relative mRNA levels of Tgfbr2, Smad3 in mouse livers. Data are presented as the mean ± SEM. n=10, *P<0.05, ** P<0.01, ***P<0.001 vs. Control.

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Staining

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: miR-552-3p overexpressed by AAV6 alleviates liver fibrosis and inflammation in CCl 4 -induced animal model. (A) Schematic diagram of CCl 4 -induced animal experiment. (B) The Desmin staining of liver tissues. (C) HE, Sirius red, Masson's trichrome and α-SMA staining of liver samples. Scale bar: 100 µm. (D-F) Fibrotic area per field was quantified. (G-H) ALT and AST levels in mouse serum. (I) The content of hydroxyproline in mouse livers. (J-K) Relative mRNA levels of fibrotic and inflammatory genes in mouse livers. (L-M) Protein expression levels of Collagen Ⅰ, p-Smad3 and t-Smad3 in mouse livers. (N) Relative mRNA levels of Tgfbr2 and Smad3 in mouse livers. Data are presented as the mean ± SEM. n=10, *P<0.05, ** P<0.01, ***P<0.001 vs. Control.

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Animal Model, Staining, Expressing

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: miR-552-3p inhibits fibrotic and inflammatory genes through the seed sequence binding to their 3'-UTR. (A-C) Relative mRNA levels of fibrotic and inflammatory genes in LX-2 cells transfected with miR-552-3p and its 3 mutants (1 nM) for 48 h. (D-G) The protein levels of Collagen I, p-Smad3 and t-Smad3 in TGF-β1/Smad3 signaling pathway in LX-2 cells transfected with miR-552-3p and its 3 mutants (1 nM) for 72 h. (H-L) The activity of luciferase in HEK293 cells transfected with 3'-UTR reporter of COL3A1, MMP-2, CCL2, SMAD3 and TGFBR2 (psiCHECK2-COL3A1/MMP-2/CCL2/SMAD3/TGFBR2) and miR-552-3p or the corresponding NC (30, 10, 1 nM). Data are presented as the mean ± SD of three independent experiments. *P<0.05, ** P<0.01, ***P<0.001 vs . NC; #P<0.05, ##P <0.01, ###P <0.001 vs . miR-552-3p; ns, non-significant.

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Sequencing, Binding Assay, Transfection, Activity Assay, Luciferase

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: Sequence alignment of miR-552-3p and the 3'-UTR of ACTA2, COL1A1, COL3A1, MMP-2, TIMP-2, SMAD3, TGFBR2, IL-6 and CCL2

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Sequencing

Journal: International Journal of Biological Sciences

Article Title: MiR-552-3p Regulates Multiple Fibrotic and Inflammatory genes Concurrently in Hepatic Stellate Cells Improving NASH-associated Phenotypes

doi: 10.7150/ijbs.80760

Figure Lengend Snippet: Schematic representation of the mechanism by which miR-552-3p alleviates liver fibrosis and inflammation. MiR-552-3p alleviates liver fibrosis and inflammation via inactivating hepatic stellate cells. miR-552-3p targets the 3'-UTRs of TGFBR2 and SMAD3 to inhibit TGF-β1/Smad3 pathway and decrease the expression of fibrotic genes indirectly, and targets to the 3'-UTRs of multiple fibrotic and inflammatory genes to decrease their expression directly, thus suppressing HSC activation and proliferation.

Article Snippet: The antibodies used were as follows: Collagen I and p-Smad3 (abcam, USA, ab260043, ab52903), t-Smad3 and GAPDH (Cell Signaling Technology, USA, 9523, 5174), α-SMA (Proteintech, 23081-1-AP).

Techniques: Expressing, Activation Assay

Journal: Journal of Veterinary Medicine

Article Title: Single-Cell Phosphospecific Flow Cytometric Analysis of Canine and Murine Adipose-Derived Stem Cells

doi: 10.1155/2017/5701016

Figure Lengend Snippet: Antibodies used in this study.

Article Snippet: Phosphorylated Human Smad2 (pS465/pS467)/Smad3 (pS423/pS425) Peptide , Smad2/3 , Mouse IgG1, κ , Alexa Fluor 647 , BD Biosciences , O72-670 , 1 : 200.

Techniques:

Journal: Journal of Veterinary Medicine

Article Title: Single-Cell Phosphospecific Flow Cytometric Analysis of Canine and Murine Adipose-Derived Stem Cells

doi: 10.1155/2017/5701016

Figure Lengend Snippet: Comparison of protein phosphorylation of mADSCs and cADSCs. The red peaks represent the isotype controls, and the blue peaks represent antigens. β -catenin, β -catenin (pS45), Akt (pS473), Stat3, Stat4, Stat5, and ERK1/2 were detected, whereas Akt (pT308), CD140b, Stat1, Stat6, p38 MAPK, and Smad2/3 were not detected in mADSCs. In addition, β -catenin, β -catenin (pS45), Akt (pS473), and Stat4 were detected, whereas Akt (pT308), CD140b, Stat1, Stat3, Stat5, Stat6, p38 MAPK, Smad2/3, and ERK1/2 were negative in cADSCs. The percentages shown in the figure represent the positive rates of the antigen.

Article Snippet: Phosphorylated Human Smad2 (pS465/pS467)/Smad3 (pS423/pS425) Peptide , Smad2/3 , Mouse IgG1, κ , Alexa Fluor 647 , BD Biosciences , O72-670 , 1 : 200.

Techniques: