Journal: Cell Death Discovery

Article Title: NR4A1 enhances MKP7 expression to diminish JNK activation induced by ROS or ER-stress in pancreatic β cells for surviving

doi: 10.1038/s41420-021-00521-0

Figure Lengend Snippet: A ROS may result in ER-stress. ROS or/and ER-stress incurs JNK activation. The activated JNK leads to pancreatic β cell apoptosis via mitochondria-dependent or independent pathways. B Acute ROS or ER-stress results in the expression of NR4A1, which in turn enhances the expression of MKP7, while MKP7 is able to abolish partial JNK activation by exerting its phosphatase activity, therefore, largely reduces pancreatic β cell apoptosis. The fate of pancreatic β cells depends on the balance between the availability of NR4A1 or MKP7 and the severity of ROS or ER-stress.

Article Snippet: The mouse pancreatic β cell lines (MIN6 and β-TC6) were from ATCC.

Techniques: Activation Assay, Expressing, Activity Assay

Journal: PLoS ONE

Article Title: Modified human glucagon-like peptide-1 (GLP-1) produced in E . coli has a long-acting therapeutic effect in type 2 diabetic mice

doi: 10.1371/journal.pone.0181939

Figure Lengend Snippet: MIN6 cells were cultured to a density of 6×10 4 cells/mL, then seeded into a 96-well flat plate (200 μL per well) and re-cultured for 24 hours. Control: cells treated with Dulbecco's phosphate buffered saline solution; Trypsin: cells treated with 0.25% trypsin; Trypsin + 6×mGLP-1: 10 μg/mL 6×mGLP-1 digested by 0.25% trypsin; GLP-1 standard: chemically synthesized native human GLP-1; Trypsin + GLP-1: 10 μg/mL GLP-1 digested by 0.25% trypsin. Cell density, measured as optical density at A490, was determined 48 h after the addition of the test compounds. The data represent the mean ± standard deviation (n = 6). The experiment was repeated three times and the results were statistically analyzed using one-way ANOVA method following Tukey post-hoc hypothesis test. The symbols a, b, and c refer to the significance level, and different symbol means statistical significance (at 5%, 1%, and 0.1% level) between treatments, or else, no significance.

Article Snippet: Based on the method described by Brandsma et al . [ ], with minor modifications, a mouse pancreatic tumor β cell line (MIN6) (iCell Bioscience Inc, Shanghai, China) was cultured and maintained in Roswell Park Memorial Institute (RPMI) 1640 medium containing 10% (v/v) fetal bovine serum, 100 U/mL penicillin, 0.1 mg/mL streptomycin and 50 μmol/L 2-mercaptoethanol at 37°C in an incubator with 5% CO 2 and 95% humidified air.

Techniques: Cell Culture, Control, Saline, Synthesized, Standard Deviation

Journal: Communications Chemistry

Article Title: A duo of redox-sensitive pore-loop cysteines controls the activity of the neural ion channel TRPM3

doi: 10.1038/s42004-026-01889-9

Figure Lengend Snippet: A Fluorescent ratio increases of non-transfected HEK293T cells (Empty HEK; green) or HEK293T cells stably expressing TRPM3α2 cells upon increasing temperatures. Cells were either non-treated (NT; black) with (open square) or without (filled square) the addition of 10 µM PS, or DTT-pretreated (for ≥ 40 min with 10 mM; yellow) with (open diamond) or without (filled diamond) the addition of 10 µM PS. Fluo4 signals were normalized to the baseline values at 16°C (ΔF/F 16°C ). B CIM0216 dose-dependency of TRPM3 in non-treated (black) and DTT-pretreated (yellow) condition. Data is represented as mean ± SEM. HEK-TRPM3 cells were tested in a 96-well plate-based assay in the presence and absence of DTT and stimulated with different doses of CIM0216. C Time course of whole-cell currents at ±80 mV recorded in non-treated (NT) HEK293T cells stably expressing TRPM3α2 upon application of the TRPM3 activator CIM0216 (1 µM). D Same as in A) but for HEK293T cells stably expressing TRPM3α2 that were pre-treated ≥40 min with the reductive agent DTT (10 mM). E Current density changes for non-treated and DTT-pre-treated HEK293T TRPM3α2 cells upon application of CIM0216 as shown in ( A ) and ( B ). Statistical comparisons performed with Student’s two-sample t -test (at +80 mV: p = 0.012, t -value = 4.15, DF = 4.40; at −80 mV: p = 0.005, t -value = −5.34, DF = 4.35). F Ratio of currents at +80 mV versus currents at −80 mV for non-treated and DTT-pre-treated HEK293T TRPM3α2 cells upon application of CIM0216. Statistical comparisons performed with a Mann-Whitney test ( p = 0.014, U = 1, Z = −2.46).

Article Snippet: To obtain homogenously shaped pancreatic β-cell spheroids, MIN6 cells were seeded in glass-bottom 96-well plates (P96-1.5H-N, Cellvis Inc.) containing 19 micro-pillar structures per well, as described previously .

Techniques: Transfection, Stable Transfection, Expressing, MANN-WHITNEY