3' mRNA-seq of human Ma-Mel47 (melanoma) and Panc89 (PDAC) cells treated with the USP7 degraders NK250 and NK266, respectively, for 72 h.
3' mRNA-seq of human Ma-Mel47 (melanoma) and Panc89 (PDAC) cells treated with the USP7 degraders NK250 and NK266, respectively, for 72 h.
The melanoma cell line Ma-Mel-47 was exposed to 1 µM NK266 and the PDAC cell line Panc89 to 1 µM NK250 against DMSO as a vehicle control in each case for 72 hours. Following treatment, cells were lysed and total RNA was isolated using the AllPrep DNA/RNA Kit (Qiagen) in accordance with the manufacturer's recommendations. RNA quality and integrity were evaluated using an Agilent TapeStation system. Library preparation was carried out following the QuantSeq 3' mRNA Library Prep Kit protocol (Lexogen). For each sample, 500 ng of RNA was used as input, and 15–20 PCR amplification cycles were performed, with the precise cycle number determined individually by qPCR. Library size distribution and concentrations were assessed on the Agilent TapeStation.