Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Cell growth and phosphorylation status of AKT in acid-tolerable H-2452AcT cells. (A, B) H-2452 and H-2452AcT cells were incubated with the RPMI-1640 medium containing (a) or not containing (b) 3.8 μM of lactic acid for 24 h, 48 h, and 72 h. The cell viability and p-AKT level were determined using an MTT assay and a western-blot analysis, respectively. (C) Cells were incubated with the RPMI-1640 medium without lactic acid for 24 h, 48 h, and 72 h. The cell distributions in the sub-G 0 /G 1 , G 0 /G 1 , S, and G 2 /M phases were analyzed using flow cytometry following a propidium-iodide staining (20 μg/ml). The error bars indicate the mean ± standard deviation for three independent experiments. The β-actin was used as a loading control. * P < .05 vs. the respective H-2452 controls.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Phospho-proteomics, Incubation, MTT Assay, Western Blot, Flow Cytometry, Staining, Standard Deviation, Control

Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Effects of cariporide and LY294002 on the cell growth and phosphorylation status of AKT in H-2452 and H-2452AcT cells. (A, B) The cells were incubated with the vehicle (0.1% DMSO) or various concentrations of cariporide (40 μM to 360 μM) alone (a) or LY294002 (2.5 μM to 20 μM) alone (b) for 48 h. (C) Cells were treated with cariporide (160 μM) and LY294002 (5 μM), alone or in combination, for 24 h, 48 h and 72 h. The cell viability was determined using an MTT assay. The p-AKT levels were determined by the western-blot analysis. The error bars indicate the mean ± standard deviation for three independent experiments. The β-actin was used as a loading control. * P < .05 vs. the respective H-2452 controls. Car, cariporide; LY, LY294002; Car/LY, the combination treatment of cariporide and LY294002.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Phospho-proteomics, Incubation, MTT Assay, Western Blot, Standard Deviation, Control

Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Effects of cariporide and LY294002 on the levels of p53 and apoptosis-regulating proteins in H-2452 and H-2452AcT cells. (A, B) The cells were incubated with the vehicle (0.1% DMSO) or various concentrations of cariporide (40 μM to 320 μM) and LY294002 (2.5 μM to 10 μM), alone (a) or in combination (160 μM cariporide and 5 μM LY294002) (b) for 48 h. The protein levels were determined by the Western-blot analysis. The p53/Bcl-2 expression ratio and relative density of protein bands were obtained from densitometric analysis of the Western blot images normalized to β-actin. Representative results are presented from one of three independent experiments; β-actin was used as a loading control. (C) The cells were transfected with 10 nM p53-targeting siRNA (sip53) or Stealth RNAi control siRNA (siC) for 24 h, 48 h and 72 h. The cell viability and p-AKT level were determined using an MTT assay and a Western-blot analysis, respectively. The error bars indicate the mean ± standard deviation for three independent experiments. The β-actin was used as a loading control. * P < .05 vs. the respective H-2452 controls. Bcl-2, B-cell lymphoma 2; PARP, poly (ADP-ribose) polymerase. Car, cariporide; LY, LY294002; Car/LY, the combination treatment of cariporide and LY294002.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Incubation, Western Blot, Expressing, Control, Transfection, MTT Assay, Standard Deviation

Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Apoptosis-promoting effects of cariporide and LY294002 in H-2452 and H-2452AcT cells. The cells were treated with cariporide (160 μM) and LY294002 (5 μM), alone or in combination, for 72 h. (A) Nuclear morphology was assessed by nuclear staining with DAPI (magnification ×40). (B) The number of apoptotic cells following annexin V-PE staining was analyzed using a Muse Cell Analyzer. Representative results are presented from one of three independent experiments. Error bars indicate the mean ± standard deviation for three independent experiments. * P < .05 vs. the respective H-2452 controls. Car, cariporide; LY, LY294002; Car/LY, the combination treatment of cariporide and LY294002; PE, phycoerythrin.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Staining, Standard Deviation

Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Effects of cariporide and LY294002 on the cell cycle and DNA damage in H-2452 and H-2452AcT cells. The cells were treated with cariporide (160 μM) and LY294002 (5 μM), alone or in combination, for 72 h. (A) The cell distributions in the sub-G 0 /G 1 , G 0 /G 1 , S, and G 2 /M phases were analyzed using flow cytometry following a propidium-iodide staining (20 μg/ml). (B) Expression levels of cell cycle-regulating molecules were measured by Western blot analysis. (C) The alkaline comet assay was conducted to assess DNA damage. The tail moment were scored per slide using the fluorescence microscope with the Comet Assay IV v4.3 software. (magnification ×40). (D) Expression levels of DNA damage-sensing molecules were measured by Western blot analysis. The β-actin was used as a loading control. Error bars indicate the mean ± standard deviation for three independent experiments. * P < .05 vs. the respective H-2452 controls. Car, cariporide; LY, LY294002; Car/LY, the combination treatment of cariporide and LY294002; p-cdc2, phosphorylated cyclin dependent kinase 2.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Flow Cytometry, Staining, Expressing, Western Blot, Alkaline Single Cell Gel Electrophoresis, Fluorescence, Microscopy, Single Cell Gel Electrophoresis, Software, Control, Standard Deviation

Journal: Molecules and Cells

Article Title: Cariporide Enhances the DNA Damage and Apoptosis in Acid-tolerable Malignant Mesothelioma H-2452 Cells

doi: 10.14348/molcells.2017.0059

Figure Lengend Snippet: Effects of cariporide and LY294002 on ROS generation and mitochondrial membrane potential (ΔΨm) in H-2452 and H-2452AcT cells. The cells were treated with cariporide (160 μM) and LY294002 (5 μM), alone or in combination, for 72 h. (A) The levels of cellular ROS were measured using flow cytometry following staining with DCF-DA (10 μM). A shift in DCF fluorescence to the right indicated an increase in ROS (reactive oxygen species) levels. (B) ΔΨm was measured by staining the cells with Rhodamine 123 (30 nM). The error bars indicate the mean ± standard deviation for the three independent experiments. * P < .05 vs. the respective H-2452 controls. DCF-DA, 2′,7′-dichlorodihydrofluorescein diacetate; Car, cariporide; LY, LY294002; Car/LY, the combination treatment of cariporide and LY294002.

Article Snippet: The H-2452 cells were maintained at 37°C in an RPMI-1640 medium (SH30027.01; GE Healthcare Life Sci., Australia) supplemented with 10% fetal bovine serum (SH30084.03; GE Healthcare Life Sci.), 100 U/ml penicillin, and 100 μg/ml streptomycin.

Techniques: Membrane, Flow Cytometry, Staining, Fluorescence, Standard Deviation

Journal: Cell Reports Medicine

Article Title: Mediator complex subunit 1 architects a tumorigenic Treg cell program independent of inflammation

doi: 10.1016/j.xcrm.2024.101441

Figure Lengend Snippet:

Article Snippet: CD8 + T cells were stained with Cell Trace Violet (Thermofisher Scientific #C34557) and then 2.5x10 4 cells were plated per well in RPMI (Cytiva #SH30255.02) in a 96 well plate with 7.5x10 4 CD3/CD28 activation beads (Miltenyi Biotec #130-095-925), 10% fetal bovine serum (FBS) (R&D Systems #S11150), and 30 IU/mL IL2 (PeproTech #212-12).

Techniques: Blocking Assay, Control, Virus, Recombinant, Activation Assay, Adjuvant, Selection, Isolation, Staining, Software

Journal: Cell Host & Microbe

Article Title: Memory profiles distinguish cross-reactive and virus-specific T cell immunity to mpox

doi: 10.1016/j.chom.2023.04.015

Figure Lengend Snippet:

Article Snippet: RPMI-1640 without L-Glutamine , Cytiva , Cat# SH30096.01.

Techniques: Recombinant, Staining, Software