Journal: Natural Products and Bioprospecting

Article Title: Scutellarin ameliorates diabetic nephropathy via TGF-β1 signaling pathway

doi: 10.1007/s13659-024-00446-y

Figure Lengend Snippet: Scutellarin Restored Podocyte Injury of the DN Mice. a Representative images of immunohistochemistry for NPHS1 and NPHS2 of the mice treated with vehicle, scutellarin or empagliflozin (× 200; scale bar = 50 µm). b Representative images of Western-blotting for NPHS1, NPHS2. c Quantitative plot of the expression of NPHS1 of the mice. d Quantitatification of NPHS1 expression of the mice. e Representative images of Western-blotting for β-catenin, Axin2, snail and DKK1 of the mice. f – i Quantifications of the protein levels for β-catenin, Axin2, snail and DKK1 from E. All data are presented as the mean ± S.D.; n = 4–6 for each group, “n” stands for the number of animals; p vs. the model group (STZ)

Article Snippet: Scutellarin (Yunnan Phytopharmaceutical Co., LTD., China); Empagliflozin (Cat. C14295412, Macklin Biochemical, China), Streptozotocin (Cat. S8050, Solarbio, China); goat anti-rabbit immunoglobulin G (IgG) horseradish peroxidase (HRP)-linked antibody (Cat. AS014, Abclonal, China); anti-mouse IgG HRP-linked antibody (Cat. 7076S, Cell Signaling Technology, USA); Methenamine Silver Plating Stain Kit (Cat. G1790, Solarbio); Glycogen Periodic Acid Schiff (PAS) Stain Kit (Cat. G1281, Solarbio); Masson’s Trichrome Stain Kit (Cat. G1340, Solarbio); Mouse MAU enzyme-linked immunosorbent assay (ELISA) Kit (Cat. JL20493, JONIN, China); Anti-DKK1 antibody (Cat. A00632, Boster Biological Technology, China); Anti-SNAIl antibody (Cat. BP0449, Boster Biological Technology); Anti-α-SMA antibody (Cat. BM0002, Boster Biological Technology); Anti-TGFB1 antibody (Cat. BA0290, Boster Biological Technology); Anti-NPHS1 antibody (Cat. BA1669, Boster Biological Technology); Anti-NPHS2 antibody (Cat. BA1688, Boster Biological Technology); Anti-SMAD2/3 antibody (Cat. BA1395, Boster Biological Technology); Anti-Phospho-SMAD2(s250) antibody (Cat. BM4693, Boster Biological Technology); Anti-FN1 antibody (Cat. BA1772, Boster Biological Technology); AXIN2 antibody (Cat. Ab32197, Abcam, USA); Anti-Phospho-SMAD3(ser425) antibody (Cat. AF3362, Affinity Biosciences, China); Anti-COL3A1 antibody (Cat. M00788, Boster Biological Technology); Anti-extracellular signal-regulated kinase (ERK)1/2 antibody (Cat. ET1601-29, HUABIO, China); Anti-ERK1(PT202/PY204) + ERK2(PT185/PY187) antibody (Cat. ET1610-13, HUABIO); Anti-P38 antibody (Cat. ET1602-26, HUABIO); Anti-Phospho-P38(Thr180 + Tyr182) antibody (Cat. ER1903-01, HUABIO); Anti-β-Actin antibody (Cat. EM21002, HUABIO); Anti-SMAD4 antibody (Cat. A5657, Abclonal); Anti-β-catenin antibody (Cat. 610154, BD Biosciences, China); Creatinine (Cr) Assay kit (Cat. C011-2-1, Nanjing Jiancheng Bioengineering Institute, China); Glucose Assay Kit (Cat. F006-1-1, Nanjing Jiancheng Bioengineering Institute); microalbunminuria (MAU) ELISA kit (Cat. JL20493, JONIN).

Techniques: Immunohistochemistry, Western Blot, Expressing

Journal: Hypertension

Article Title: Single-Nucleotide Polymorphisms of the Dopamine D2 Receptor Increase Inflammation and Fibrosis in Human Renal Proximal Tubule Cells

doi: 10.1161/hypertensionaha.113.02569

Figure Lengend Snippet: Figure 4. SMAD3 and Snail1 expression in renal proximal tubular cells (RPTCs) carrying dopamine D2 receptor (D2R) wild type (WT) or D2R single-nucleotide polymorphisms (SNPs). A, Expressions of SMAD3 and SNAIL1 mRNA in RPTCs from subjects carrying D2R WT or D2R SNPs were quantified by quantitative real- time polymerase chain reaction; GAPDH mRNA was used for normalization of the data, n=5 per group. *P<0.05. B, Laser- scanning confocal images of immunofluorescence staining of Snail1 in RPTCs carrying D2R WT or D2R SNPs. DAPI was used to immunostain the nuclei. Immunostaining was performed in all 10 cell lines (5 WTs and 5 bearing the SNPs). Thirty to fifty cells were observed in each sample.

Article Snippet: Snail1 was visualized with an anti-human Snail1 antibody conjugated with an NL557 fluorochrome (R&D Systems).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Immunostaining

Journal: Hypertension

Article Title: Single-Nucleotide Polymorphisms of the Dopamine D2 Receptor Increase Inflammation and Fibrosis in Human Renal Proximal Tubule Cells

doi: 10.1161/hypertensionaha.113.02569

Figure Lengend Snippet: Figure 7. Transforming growth factor-β (TGF-β) and Snail1 expression in renal proximal tubular cells (RPTCs) from subjects carrying dopamine D2 receptor (D2R) single-nucleotide polymorphisms (SNPs) transfected with DRD2 wild-type (WT) plasmid or empty vector. A, Expressions of TGF-β and Snail1 mRNA in RPTCs from subjects carrying D2R SNPs transfected with DRD2 WT plasmid or empty vector were quantified by quantitative real-time polymerase chain reaction; GAPDH mRNA was used normalization of the data, n=5 per group. *P<0.05, t test. B, RPTC homogenates were immunoblotted using TGF-β– specific antibody; GAPDH protein was used for normalization of the data, n=5. *P<0.05, t test. C and D, Laser-scanning confocal images of immunofluorescence staining of TGF-β and Snail1 in RPTCs from subjects carrying D2R SNPs transfected with DRD2 WT plasmid or empty vector. DAPI was used to immunostain the nuclei. Immunostaining was performed in 5 cell lines carrying D2R SNPs transfected with DRD2 WT plasmid and the same cell lines transfected with empty vector. Thirty to fifty cells were observed in each sample.

Article Snippet: Snail1 was visualized with an anti-human Snail1 antibody conjugated with an NL557 fluorochrome (R&D Systems).

Techniques: Expressing, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Immunofluorescence, Staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Comparison of FNDC5/Ir (( A )—IRS score 8) expression with E-cadherin (( B )—IRS score 12), N-cadherin (( C )—IRS score 8), SNAIL (( D )—IRS score 3 and % of nuclear expression 4, ( E )—IRS score 12 and % of nuclear expression 4), SLUG (( F )—IRS score 8 and % of nuclear expression 3) and TWIST (( G )—IRS score 4 and % of nuclear expression 2, ( H )—IRS score 4 and % of nuclear expression 2) using immunohistochemistry (IHC) (positive reactions—brown cell cytoplasm) in breast cancer (BC), magnification ×200.

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemistry

Journal: International Journal of Molecular Sciences

Article Title: The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer

doi: 10.3390/ijms24108628

Figure Lengend Snippet: Correlation of FNDC5/Ir expression level with E-cadherin ( A ) and N-cadherin ( B ), cytoplasmic ( C ) and nuclear ( D ) SNAIL expression levels, cytoplasmic ( E ) and nuclear ( F ) SLUG expression levels, cytoplasmic ( G ) and nuclear ( H ) TWIST expression levels in breast cancer (BC) (sample size n = 541).

Article Snippet: The expression of proteins was detected by specific primary antibodies, i.e., polyclonal rabbit: anti-irisin/FNDC5 (dilution 1:50; code no. NBP2-14024; Novus Biologicals, Littleton, CO, USA) and anti-SNAIL (1:400, Clone, code 13099-1-AP, Proteintech, Rosemont, IL, USA), monoclonal mouse: anti-E-cadherin antibody (ready to use, Clone NCH-38, code IR059; Dako, Glostrup, Denmark), anti-N-cadherin (1:50, Clone 6G11, code M3613; Dako), anti-SLUG (1:50, clone A-7, sc-166476, Santa Cruz Biotechnology, Santa Cruz, CA, USA), and monoclonal mouse anti-TWIST (dilution 1:50, clone Twist2C1a, code ab-50887, Abcam, Cambridge, UK).

Techniques: Expressing