Journal: International Journal of Molecular Sciences

Article Title: Novel Benzoxazoles Containing 4-Amino-Butanamide Moiety Inhibited LPS-Induced Inflammation by Modulating IL-6 or IL-1β mRNA Expression

doi: 10.3390/ijms23105331

Figure Lengend Snippet: Compounds 5f and 4d attenuate inflammation in vitro. Effects of compounds (10 µM) on the protein levels of STAT3, IκB, and NF-κB in AML-12 cells. ( a ) Expression levels of inflammation related-proteins in vitro. ( b – d ) Protein expression levels were normal-ized against the indicated protein. * p < 0.05 compared with the group treated with the PBS (vehicle). + p < 0.05 compared with the group treated with the LPS. Data are pre-sented as the mean ± SD.

Article Snippet: The human keratinocytes HaCaT or the alpha mouse liver 12 (AML-12) cells were obtained from Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: In Vitro, Expressing

Journal: Heliyon

Article Title: Oral administration of kynurenic acid delays the onset of type 2 diabetes in Goto-Kakizaki rats

doi: 10.1016/j.heliyon.2023.e17733

Figure Lengend Snippet: KYNA enhances mRNA and protein expression of hepatic UCP genes. (A, B) Real-time quantitative PCR analysis (n = 6) of gene expression in HepG2 cells after 48 h of 10 μmol/L (K10) or 100 μmol/L (K100) KYNA treatment compared with control group (white punctuated line). (C, D) Western blot analysis of UCP2 expression in HepG2 cells after 48 h of 100 μmol/L KYNA treatment (n = 3), (E, F) Real-time quantitative PCR analysis of gene expression in HepG2 cells after 2 h treatment with 10 μmol/L (K10) or 100 μmol/L (K100) KYNA (n = 6), (G) Quantitative PCR analysis of gene expression in AML-12 cells after 48 h treatment with 10 μmol/L (K10) or 100 μmol/L (K100) KYNA (n = 6), (H) Expression of hepatic energy metabolism genes in GK rats after oral administration 25 mg/kg KYNA (KYNA) or NaCl (Ctrl) (I) Comprehensive analysis of transcriptomic pathways in KYNA-treated HepG2 cells after 48 h KYNA, 100 μmol/L (n = 3). Statistical analysis using t -test, * p < 0.05.

Article Snippet: HepG2 human liver cells and AML-12 mouse liver cells (Procell, China) were cultured in MEM basal medium (Gibco, China) containing 10% fetal bovine plasma (Gibco, Australia).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Frontiers in pharmacology

Article Title: Brg1 and RUNX1 synergy in regulating TRPM4 channel in mouse cardiomyocytes.

doi: 10.3389/fphar.2024.1494205

Figure Lengend Snippet: FIGURE 8 Brg1 modulated TRPM4 promoter activity by interaction with RUNX1. (A) Predicting the residue label and hydrogen bond length of interaction between Brg1 and RUNX1, and the combination state and surface structure of Brg1 and RUNX1 by Protein Data Bank. (B) Co-immunoprecipitation assays of Brg1 and RUNX1 on the nucleoprotein extracted from neonatal mouse cardiomyocytes. (C) RUNX1 significantly boosted TRPM4 promoter activity. Statistical analysis was performed with two-tailed Student’s t-test. *p < 0.05, **p < 0.01 vs. NC group. n = 8 in each group. (D) Brg1 knockdown significantly decreased the TRPM4 promoter activity. **p < 0.01 vs. Scramble-siRNA group by two-tailed Student’s t-test. n = 6 in each group. (E) Inhibited Brg1 by PFI-3 (10 μM) significantly reduced the TRPM4 promoter activity. **p < 0.01 vs. DMSO group by two-tailed Student’s t-test. n = 6 in each group. (F) RUNX1 knockdown declined the increases in TRPM4 promoter activity caused by Brg1-OE. *p < 0.05, vs. NC group; ##p < 0.01 vs. +Scramble-siRNA by (Continued)

Article Snippet: After incubation in 5% non-fat milk for 1.5 h at room temperature, the membranes were incubated at 4°C overnight with TRPM4 antibody (1:500; ABclonal Technology Co.,Ltd.), β-actin antibody (1:1000; Santa, United States), Brg1 antibody (Sigma-Aldrich, 07–478, 1:500) and RUNX1 antibody (Proteintech 25315-1-AP, 1:1000).

Techniques: Activity Assay, Residue, Immunoprecipitation, Two Tailed Test, Knockdown

Journal: Frontiers in pharmacology

Article Title: Brg1 and RUNX1 synergy in regulating TRPM4 channel in mouse cardiomyocytes.

doi: 10.3389/fphar.2024.1494205

Figure Lengend Snippet: FIGURE 9 A schematic diagram summarizing the potential mechanisms that Brg1 interacted with RUNX1 transcriptional regulated TRPM4, that eventually leads to TRPM4 overactivation in cardiomyocytes induced by hypoxia.

Article Snippet: After incubation in 5% non-fat milk for 1.5 h at room temperature, the membranes were incubated at 4°C overnight with TRPM4 antibody (1:500; ABclonal Technology Co.,Ltd.), β-actin antibody (1:1000; Santa, United States), Brg1 antibody (Sigma-Aldrich, 07–478, 1:500) and RUNX1 antibody (Proteintech 25315-1-AP, 1:1000).

Techniques:

Journal: International journal of molecular medicine

Article Title: Expression of PADI4 in patients with ankylosing spondylitis and its role in mediating the effects of TNF-α on the proliferation and osteogenic differentiation of human mesenchymal stem cells.

doi: 10.3892/ijmm.2015.2248

Figure Lengend Snippet: Figure 8. Western blot analysis of bone morphogenetic protein 2 (BMP-2), runt- related transcription factor 2 (Runx) and Osterix protein expression in human mesenchymal stem cells (hMSCs) following transfection with peptidyl arginine deiminase, type IV (PADI4) siRNA in the presence or absence of tumor necrosis factor-α (TNF-α). (A) Blots showing BMP-2, Runx2, Osterix and β-actin protein expression. Lane 1, vehicle-treated group; lane 2, vehicle + PADI4 siRNA-treated group; lane 3, TNF-α-treated group; lane 4, TNF-α + PADI4 siRNA-treated group. (B) Relative protein expression of BMP-2. (C) Relative protein expression of Runx2. (D) Relative protein expression of Osterix. β-actin was used as a loading control. *P<0.05 compared with the vehicle-treated group. **P<0.05 compared with the TNF-α-treated group.

Article Snippet: Following blocking with 5% non-fat milk, the membranes were incubated with anti-PADI4 rabbit polyclonal antibody (Cat. no. sc-98991, 1:500 dilution; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-bone morphogenetic protein 2 (BMP-2) mouse monoclonal antibody (Cat. no. ab6285, 1:400 dilution), anti runt-related transcription factor 2 (Runx2) rabbit polyclonal antibody (Cat. no. ab102711, 1:400 dilution), anti-Osterix mouse monoclonal antibody (Cat. no. ab57335, 1:800 dilution) (all from Abcam, Cambridge, MA, USA) and anti β-actin mouse monoclonal antibody (Cat. no. BM0627, 1:1,000 dilution; Boster, Wuhan, China) at 37 ̊C for 2 h. The membranes were washed 3 times with TBST and incubated with rabbitanti mouse IgG (Cat. no. sc-358913, 1:2,000 dilution) or mouse-anti rabbit IgG (Cat. no. sc-2357, 1:2,000 dilution) horseradish peroxidase (HRP)-conjugated secondary antibody (both from Santa Cruz Biotechnology) at 37 ̊C for 1 h. The signals were detected using an ECL detection kit (Pierce Biotechnology, Inc.).

Techniques: Western Blot, Expressing, Transfection, Control