Journal: Experimental and Therapeutic Medicine

Article Title: Tilianin attenuates MPP + -induced oxidative stress and apoptosis of dopaminergic neurons in a cellular model of Parkinson's disease

doi: 10.3892/etm.2022.11223

Figure Lengend Snippet: Effects of tilianin on the viability of MES23.5 cells. (A) Chemical structure of tilianin. (B) Effects of tilianin at different concentrations on cell viability were examined using the Cell Counting Kit-8 assay. Results are presented as the mean ± SD.

Article Snippet: The dopaminergic neuron MES23.5 cell line (cat. no. CVCL-J351; http://www.biovector.net/product/2277098.html ) was acquired from the BioVector National Type Culture Collection, Inc. MES23.5 cells were maintained in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Merck KGaA), 2% Sato's solution containing 25 mg insulin, transferrin, selenium and sodium pyruvate solution (ITS-A; cat. no. 51300044; Thermo Fisher Scientific, Inc.), 0.315 mg/ml progesterone (cat. no. HY-N0437; MedChemExpress) and 20 mg putrescine (cat. no. HY-N2407; MedChemExpress), in addition to 100 U/ml penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37˚C.

Techniques: Cell Counting

Journal: Experimental and Therapeutic Medicine

Article Title: Tilianin attenuates MPP + -induced oxidative stress and apoptosis of dopaminergic neurons in a cellular model of Parkinson's disease

doi: 10.3892/etm.2022.11223

Figure Lengend Snippet: Effects of tilianin on the MAPK pathway in MPP + -stimulated MES23.5 cells. The expression levels of p-p38MAPK, p38MAPK, p-ERK1/2, ERK1/2, p-JNK and JNK in cells stimulated with MPP + with or without tilianin preconditioning were detected by western blotting. Results are presented as the mean ± SD. ** P<0.01, *** P<0.001. MPP + , 1-methyl-4-phenylpyridinium; p, phosphorylated.

Article Snippet: The dopaminergic neuron MES23.5 cell line (cat. no. CVCL-J351; http://www.biovector.net/product/2277098.html ) was acquired from the BioVector National Type Culture Collection, Inc. MES23.5 cells were maintained in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Merck KGaA), 2% Sato's solution containing 25 mg insulin, transferrin, selenium and sodium pyruvate solution (ITS-A; cat. no. 51300044; Thermo Fisher Scientific, Inc.), 0.315 mg/ml progesterone (cat. no. HY-N0437; MedChemExpress) and 20 mg putrescine (cat. no. HY-N2407; MedChemExpress), in addition to 100 U/ml penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37˚C.

Techniques: Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Heat shock promotes inclusion body formation of mutant huntingtin (mHtt) and alleviates mHtt-induced transcription factor dysfunction

doi: 10.1074/jbc.RA118.002933

Figure Lengend Snippet: Sorbitol, a chemical chaperone, promoted the structuring of diffusible assemblies of mHtt to form IBs and supported survival of the E14 rat striatal neuron–derived ST14A cell line. ST14A cells were transfected with the HttQ74–EGFP plasmid DNA and then plated in 96-well plate (day 0) according to methods described in the text. Sorbitol was added to designated wells of cells to a final concentration of 150 mm at 24 h after plating (day 1). A–D, for cell imaging, cells were fixed 72 h after plating (day 3) and processed to stain for HSP70 and nuclei according to methods described in the text. A and B are images of the control and C and D are of sorbitol-treated (150 mm, 48 h) cells. These images are representative of results from two separate experiments each with eight independent samples. E, to test for survival of the control and sorbitol-treated cells under stress, the culture medium was changed to DMEM without serum and cells were moved from a 33 °C to a 39 °C incubator on day 2 after plating. Cell viability was determined 2 days after the shift to this restrictive condition using the CellTiter-Glo reagent from Promega (G7572). Result represents the average of viability readouts (3X each sample) of 16 independent samples per condition from two separate experiments. F, the time line of the experiment is as shown. Probability of difference <0.001 is extremely significant (***). Error bars represent S.D.

Article Snippet: Culturing of the ST14A rat E14 striatal neuron–derived cell line was done according to instructions provided by the HD Community Biorepository via Coriell Institute for Medical Research in DMEM with 10% fetal bovine serum.

Techniques: Derivative Assay, Transfection, Plasmid Preparation, Concentration Assay, Imaging, Staining, Control