HEK 293T cells were plated in six-well culture plates at 5.0 × 105 cells/well in DMEM. The next day, the indicated plasmids were mixed with an empty vector to a total of 2.0 μg DNA in 125 μl in Opti-MEM and transfected using FuGENE HD (Promega) according to the manufacturer’s protocol. Unless indicated otherwise, 0.02 μg CARD8, 0.02 μg NLRP1, and 0.005 μg ASC were used. The next day, the cells were treated as described. For microscopy experiments, the cells were plated directly into Nunc LabTek II Chamber slide w/Cover sterile glass slides (154534; Thermo Fisher Scientific) at 8.0 × 104 cells/well and treated with 25 μl transfection master mix dropwise.
Nunc lab tek 2 chamber slide w cover sterile glass slides
Manufactured by Thermo Fisher Scientific
The Nunc Lab-Tek II Chamber slide w/Cover sterile glass slides are a laboratory consumable product designed for cell culture and microscopy applications. The product consists of sterile glass slides with attached chambers that allow for the controlled culturing and observation of cells. The core function of this product is to provide a designated, contained environment for cell growth and analysis.
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3 protocols using nunc lab tek 2 chamber slide w cover sterile glass slides
1
Transfection of HEK 293T Cells
2
CARD8 Plasmid Transfection in HEK293T Cells
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HEK293T cells were plated in 6-well culture plates at 5.0 × 105 cells/well in DMEM. The next day, the indicated plasmids were mixed with an empty vector to a total of 2.0 μg DNA in 125 μL Opti-MEM and transfected using FuGENE HD (Promega) according to the manufacturer’s protocol. Unless indicated otherwise, 0.02 μg CARD8 (or CARD8 chimera) was used. After 16-20 h, the cells were treated as described. For microscopy experiments, cells were plated directly into Nunc Lab-Tek II Chamber slide w/Cover sterile glass slides (Thermo Fisher Scientific, 154534) at 2.5 × 104 cells/well and treated with 25 μL transfection master mix dropwise.
3
CARD8 Plasmid Transfection in HEK293T Cells
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