ARPE-19 cells (ATCC, CRL-2302) were cultured in DMEM/F12 (Cat. No: 11320033; Thermo Fisher, Darmstadt, Germany) supplemented with 10% FBS (Cat. No: 35-015-CF, Corning, Corning, NY, USA) and 1% Streptomycin (Cat. No: BS.A 2213, Bio&SELL, Feucht / Nürnberg, Germany) at 37°C and 5% CO2.
The day before the experiment, semi confluent cells were cultured in serum-free medium. The cells were incubated with 0.03% saccharin (Cat. No: 223370010; Thermo Fisher, Darmstadt, Germany) in serum-free medium for 24 hours. RNA-isolation was performed using the RNeasy Plus Mini Kit (Cat. No: 74136; Qiagen, Hilden, Germany), followed by reverse transcription using the QuantiTect Reverse Transcription Kit (Cat. No: 205311; Qiagen, Hilden, Germany). Primers for qPCR were obtained from Eurofins Genomics (Ebersberg, Germany; seeSupplementary Table S2 ). The qPCR was conducted with the Biozym SYBR green PCR kit (Cat. No: 331416S; Biozym, Hess, Oldendorf, Germany) on a RotorGene (Qiagen, Hilden, Germany) and analyzed using the ddCT method. The detection of TAS1R3 expression in ARPE-19 cells was performed by PCR followed by 2% agarose gel electrophoresis with ethidium bromide.
The day before the experiment, semi confluent cells were cultured in serum-free medium. The cells were incubated with 0.03% saccharin (Cat. No: 223370010; Thermo Fisher, Darmstadt, Germany) in serum-free medium for 24 hours. RNA-isolation was performed using the RNeasy Plus Mini Kit (Cat. No: 74136; Qiagen, Hilden, Germany), followed by reverse transcription using the QuantiTect Reverse Transcription Kit (Cat. No: 205311; Qiagen, Hilden, Germany). Primers for qPCR were obtained from Eurofins Genomics (Ebersberg, Germany; see
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