PANC-1 cells were collected from a confluent flask, split 1:5 and plated on 35 mM µ-Dishes with a polymer coverslip bottom (Ibidi) and incubated in a humidified 37 °C incubator with 5% CO2 for 24 h. The next day cells were transfected with pEGFP-C3 KRAS4B 12 V or pEGFP-C3 NRAS 12D using Lipofectamine 3000 (Thermo Fisher) following manufacturer’s instructions, and incubated for 3 days in a humidified 37 °C incubator with 5% CO2. Cells in 1.8 ml PBS with 10%FCS were imaged in a Nikon Eclipse Ti inverted microscope with a humidified live cell imaging chamber using NIS-Elements software. 200 µl of PBS with 10% DMSO only or of 200 µM DW0254 previously diluted in PBS with 10% DMSO were added, and samples were imaged every 5 min for 1 h.
Dishes with a polymer coverslip bottom
Manufactured by Ibidi
The µ-Dishes with a polymer coverslip bottom are a type of lab equipment designed for cell culture applications. The dishes feature a thin, transparent polymer material at the bottom, which allows for high-quality microscopic imaging of cells. The core function of these dishes is to provide a suitable substrate for cell attachment and growth, while enabling optical clarity for observation and analysis.
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Lab products found in correlation
2 protocols using dishes with a polymer coverslip bottom
1
Real-Time Imaging of KRAS and NRAS Mutants
2
Live-cell Imaging of KRAS/NRAS Mutants
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PANC-1 cells were collected from a confluent flask, split 1:5 and plated on 35mM µ-Dishes with a polymer coverslip bottom (Ibidi) and incubated in a humidified 37°C incubator with 5% CO2 for 24 hours. The next day cells were transfected with pEGFP-C3 KRAS4B 12V or pEGFP-C3 NRAS 12D using Lipofectamine 3000 (Thermo Fisher) following manufacturer's instructions, and incubated for 3 days in a humidified 37°C incubator with 5% CO2. Cells in 1.8ml PBS with 10%FCS were imaged in a Nikon Eclipse Ti inverted microscope with a humidified live cell imaging chamber using NIS-Elements software. 200µl of PBS with 10% DMSO only or of 200µM DW0254 previously diluted in PBS with 10% DMSO were added, and samples were imaged every 5 minutes for 1 hour.
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