Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibited ox-LDL-induced CD36 expression and Ca 2+ influx. (A) Histogram showing CD36 + macrophages in RAW264.7 macrophage after 100 µg/mL ox-LDL treatment were quantified by FCM. Statistical results of the proportion (B) and MFI (C) from (A) (n = 3). (D-E) The protein levels of CD36 by Western Blot in RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL and KAE(n = 3). (F) The fluorescence images of CD36 in different groups of RAW264.7 macrophage after treatment with 100 µg/mL ox-LDL. (G) Statistical results of fluorescence images from (F) (n = 4). (H) Representative fluorescence images of Ca 2+ in different groups of RAW264.7 macrophage (I) Statistical results of fluorescence images from (H) (n = 4). The results represent means ± SEM. *P < 0.05, **P < 0.01 was compared with the control group. # P < 0.05 was compared with the ox-LDL group.

Article Snippet: Then, to prevent nonspecific binding, tissues 191 sections or cells were blocked with 4% bovine serum albumin for 1.5 h. Then, the tissues sections or cells were treated with primary antibodies, including IL-1β antibody (1:100, Wanlei), Piezo1 antibody (1:100, Proteintech), Nrf2 (1:100, CST), CD36 (1:100, CST) for 12 h at 4 °C.

Techniques: Expressing, Western Blot, Fluorescence, Control

Journal: Journal of Advanced Research

Article Title: Kaempferol regulating macrophage foaming and atherosclerosis through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway

doi: 10.1016/j.jare.2024.11.016

Figure Lengend Snippet: KAE inhibits macrophage inflammation and through Piezo1-mediated MAPK/NF-κB and Nrf2/HO-1 signaling pathway. (A-B) The protein expression of CD36 by Western Blot in BMDMs after treatment (n = 3). (C) Representative fluorescence images of CD36 in different groups of BMDMs after treatment with 100 µg/mL ox-LDL. (D) Statistical results of fluorescence images from (C) (n = 4). (E-H) The protein expression of ERK, p-ERK, JNK, p-JNK, p38 and p-p38, by Western Blot in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). (I-J) The protein expression of p-p65, and p65 by Western Blot in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). (K-M) The protein expression of HO-1, and Nrf2 by in BMDMs after treatment with 100 µg/mL ox-LDL (n = 3). The results represent means ± SEM. *P < 0.05, **P < 0.01.

Article Snippet: Then, to prevent nonspecific binding, tissues 191 sections or cells were blocked with 4% bovine serum albumin for 1.5 h. Then, the tissues sections or cells were treated with primary antibodies, including IL-1β antibody (1:100, Wanlei), Piezo1 antibody (1:100, Proteintech), Nrf2 (1:100, CST), CD36 (1:100, CST) for 12 h at 4 °C.

Techniques: Expressing, Western Blot, Fluorescence

Journal: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: Visualization of molecular docking results using interaction diagrams. ( A ) Molecular docking of CAT and ERK1. ( B ) Molecular docking of CAT and MMP2. ( C ) Molecular docking of CAT and NOX4. ( D ) Molecular docking of CAT and RAGE. ( E ) Molecular docking of CAT and S100A8.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques:

Journal: Journal of Inflammation Research

Article Title: Catalpol Alleviates HFpEF via Inhibition of the S100A8-RAGE-NOX4 Inflammatory Axis in Murine Hearts

doi: 10.2147/JIR.S552741

Figure Lengend Snippet: CAT downregulated the S100A8-RAGE-NOX4 signaling pathway in HFpEF. ( A ) Representative WB bands of S100A8, RAGE, NOX4, ERK, p-ERK, P65, p-P65, and MMP2. ( B – G ) Quantification of protein expression levels analyzed using ImageJ software (n=3). ( H ) Quantitative analysis of IL-1β expression (n=3). ( I ) Quantitative analysis of IL-6 expression (n=3). ( J ) Quantitative analysis of TNF-α expression (n=3). Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Primary antibodies against the following targets were used: S100A8 (15792-1-AP), RAGE (16346-1-AP), NOX4 (14347-1-AP), MMP2 (10373-2-AP), and GAPDH (60004-1-AP) from Proteintech (Wuhan, China); ERK1/2 (9102), phospho-ERK1/2 (p-ERK1/2, 4370), p65 (8242), and phospho-p65 (p-p65, 3033) from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Expressing, Software

Journal: Chemical biology & drug design

Article Title: 2-Aminothiophene derivatives as a new class of positive allosteric modulators of glucagon-like peptide 1 receptor

doi: 10.1111/cbdd.14039

Figure Lengend Snippet: Potential allosteric effect of S-1 and S-2 on human GLP-1R. (a) The effect of GLP-1 on HEK293 cells expressing human GLP-1R or empty vector in the presence or absence of S-1 (23.5 μM). (b) The effect of GLP-1 on HEK293 cells expressing human GLP-1R or empty vector in the presence or absence of S-2 (71 μM). GLP-1R activation was assessed as luminescence normalized to protein concentration and plotted as luminescence fold change with respect to vehicle control (0.5% DMSO). Data are average of three independent experiments with at least three technical replicates for each conditions, and error bars for each concentration were plotted as SEM (n = 3). Statistical analysis was done using 2-way ANOVA (****p < 0.0001; **p < 0.001; *p < 0.5). The comparison is done between the data points of the dose–response curve generated due to GLP-1’s effect on GLP-1R in the presence and absence of S-1 or S-2

Article Snippet: HEK293 cell stably expressing CRE/CREB luciferase reporter gene (BPS Bioscience #60515), RPMI medium (Corning #10-040), Krebs-Ringer Solution, HEPES-buffered (Alfa Aesar #J67795-K2), L-Glutamine (Gibco #25030-081), HEPES (Gibco #15630-080), Sodium Pyruvate (Gibco #11360-070), β-mercaptoethanol (MP #806444), D-Glucose (#G-7528), Fetal Bovine Serum (Fisher #03600511), penicillin/streptomycin (Corning #30-002-Cl), Hygromycin B (Alfa Aesar #J60681), Lipofectamine 2000 (Invitrogen #11668027), vasoactive intestinal peptide receptor (VIPR) peptide agonist (Sigma #V3628), VIPR peptide antagonist (Sigma #SCP0260), GLP-1R peptide agonist (Sigma #9416), 6-well cell culture plates (Ultra Cruz #sc-204443), 96-well cell culture plates (Sigma #CLS9102), Luciferase cell culture lysis reagent (Promega #E1531), Luciferase assay reagent (Promega #E1501), and Ultra-Sensitive Rat Insulin Kit (Crystal Chem #90060) were purchased from vendors.

Techniques: Expressing, Plasmid Preparation, Activation Assay, Protein Concentration, Control, Concentration Assay, Comparison, Generated