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

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet: Global TRPM2 knockout abolishes the exacerbation of ischemic brain injury by hyperlipidemia (A) The presence of atherosclerotic plaque (indicated by black arrows) at the bifurcation of common carotid artery (CCA) and in the internal carotid artery (ICA) (scale bar size: 2 mm). (B–E) Triphenyl tetrazolium chloride (TTC) staining 24 h after MCAO (60-min) and neurological deficit score in WT, Apoe −/− mice, and Apoe −/− gM2KO mice (B and C) and in Trpm2 fl/fl mice, Apoe −/− Trpm2 fl/fl cre − mice, and Apoe −/− Trpm2 fl/fl cre + mice (D and E) fed with or without HFD ( n = 12–20 mice per group) (scale bar size: 5 mm). (F and G) TRPM2 expression in brains from WT mice fed with or without HFD ( n = 5 and 5 mice). (H and I) TRPM2 and CD36 expression in primary neurons, CECs, BMDMs, and peripheral leukocytes isolated from WT mice ( n = 5, 5, 5, and 5 dishes of cells isolated from at least 5 mice). (J and K) TRPM2 expression in peripheral leukocytes isolated from WT mice fed with HFD for 0, 1, 2, 3, and 4 months ( n = 5, 5, 5, 5, and 5 dishes of cells isolated from at least 5 mice). (L) ELISA measurement of TRPM2 levels in lysates from B cells, T cells, monocytes, and neutrophils after cell sorting of peripheral leukocytes from WT mice with or without HFD treatment. (M and N) ELISA measurement of plasma oxLDL and IL-1β levels in WT mice fed with HFD for 0, 1, 2, 3, and 4 months ( n = 5, 5, 5, 5, and 5 plasma samples from different mice). (O and P) Correlation of plasma IL-1β level with plasma oxLDL level (O) and TRPM2 expression (P) in peripheral leukocytes ( n = 5 mice). (Q) Correlation of plasma oxLDL level with TRPM2 expression in peripheral leukocytes ( n = 5 mice). Error bars: mean ± SEM; ns, no statistical significance, ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001. See also Figure S2 and Table S1 .

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Knock-Out, Staining, Expressing, Isolation, Enzyme-linked Immunosorbent Assay, FACS, Clinical Proteomics

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet: Increase of TRPM2 expression by oxLDL compromises resistance of endothelial cells to ischemia (A–C and E) TRPM2 and pp65 expression in primary CECs isolated from WT mouse treated with oxLDL (A and B) and LDL (C and E) for 0, 6, 12, 18, 24, and 48 h ( n = 5). (D and F) CD36 expression in primary CECs isolated from WT mouse treated with oxLDL for 0, 6, 12, 18, 24, and 48 h ( n = 5). (G and H) TRPM2 expression in WT mouse primary CECs treated with oxLDL for 48 h with the co-treatment of DMSO, SSO, and scavengers Mn (III) TBAP and L-NMA ( n = 5). (I and J) TRPM2, iNOS, and pp65 expression in WT and M2KO mouse primary CECs treated with oxLDL for 48 h with the co-treatment of DMSO, SN50, ACA, and TAT-M2 ( n = 5). (K and L) Examination of endothelial responses to in vitro ischemia insult. TRPM2, iNOS, pp65, and occludin expression in WT and M2KO mouse primary CECs subjected to OGD for 8 h ( n = 5). (M and N) Monitoring of the loss of giga-seal after OGD in transwell inserts plated with WT or M2KO mouse primary CECs pretreated with oxLDL for 48 h ( n = 3). (O) Measurement of the leakage of Evans blue (related to the PBS-Con groups) from upper into lower chamber 8 h after OGD in transwell inserts ( n = 6). n means X dishes of cells isolated from at least X mice per group; error bars: mean ± SEM; ns, no statistical significance, ∗, p < 0.05, ∗∗, p < 0.01, ∗∗∗, p < 0.001.

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Expressing, Isolation, In Vitro

Journal: Cell Reports Medicine

Article Title: TRPM2 overactivation drives hyperlipidemia-induced dysfunction of myeloid cells and neurovascular units

doi: 10.1016/j.xcrm.2025.101998

Figure Lengend Snippet:

Article Snippet: CD36 (D8L9T) Rabbit mAb , Cell Signaling Technology , Cat#14347; RRID: AB_2798555.

Techniques: Recombinant, Clinical Proteomics, Sequencing, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Knock-Out, Generated, Software, Microscopy, Imaging