The DLR assays were performed as described previously (38 (link), 40 (link)–42 (link), 53 (link)). In short, HEK293T cells were plated on 24 well dish (Corning) at a density of 1 × 105 cells per well-overnight before transfection. Cells were then co-transfected with 100 ng of the indicated expression plasmid, 100 ng of IFN-β or NF-κB promoter reporter and 10 ng of pRL-TK (internal control) to normalize transfection efficiency. Twenty-four hours post-transfection, the luciferase activity was detected with a luciferase assay kit (Promega).
24 well dish
Manufactured by Corning
The 24-well dish is a laboratory equipment designed to hold and cultivate cells or tissues. It features a grid of 24 individual wells, each with a consistent surface area and volume, enabling controlled and replicable experimental conditions.
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Lab products found in correlation
3 protocols using 24 well dish
1
Quantifying Cellular Signaling Pathways
2
Curcumin Treatment Impacts Cell Proliferation
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The pRc/RSV-derived eukaryotic expression vectors pRc-HA-v-Myc and pRc-v-Src have been described (34 (link)). Transcriptional transactivation analysis using the luciferase reporter system, including the firefly luciferase constructs pGL3-WS5 (pLUC-WS5) and the empty vector pGL3-Basic (pLUC), has been described (33 (link), 34 (link), 36 (link)). The eukaryotic expression construct pcDNA3.1-Rluc used for constitutive renilla luciferase expression, and the empty pcDNA3.1 vector have been described (34 (link)). Proliferation of cells treated with curcumin (CRM) was monitored in real time by using the live cell imaging system IncuCyte S3 (Essen Bioscience/Sartorius, Vienna, Austria) as described (33 (link)). Cells were seeded in a 24-well dish (Corning, Vienna, Austria), and incubated overnight. Curcumin was then added to final concentrations of 5–40 µM. Cells were monitored for 3 d by phase contrast imaging every 6 h from 16 separate regions per well using a 10× objective.
3
Wound Healing and Invasion Assays
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Cells were seeded to complete confluence in a monolayer in a 24-well plate for wound healing assay. A wound was created by scratching firmly with a 20 µl tip and baseline (time zero) images were captured. Cell were incubated at 37ºC in a humidified incubator with 5% CO 2 and photographed at the indicated time-points and the percent wound remaining was measured and plotted. Invasion assay was performed using 24-well BD BioCoat Matrigel invasion chambers (BD Biosciences, San Jose, CA) according to the manufacturer's instructions. Briefly, inserts were first rehydrated using DMEM containing 10% FBS for a final volume of 0.5 ml in the top inserts and 0.75 ml in the wells of a 24-well dish (Corning Inc., Corning, NY) for at least 2 h. Following this, the medium was removed from the upper inserts and a total of 1X10 5 cells were plated in 0.5 ml of serum free medium. Cells were allowed to invade through the matrigel towards the FBS containing medium, which acts as a chemoattractant, for 48 h. Non-invading cells were removed using a damp cotton swab while the invading cells were fixed in methanol for 10 mins and stained with 0.5% crystal violet (Sigma, St. Louis, MO) stain. Cells in at least six randomly selected fields were then counted under a light microscope.
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