Isogenic cell lines for Tsc1/Tsc2 knockout

We used murine kidney-derived principal cell lines, i.e., mIMCD3, purchased from ATCC (ATCC^® CRL-2123^TM, Manassas, VA, United States). We previously developed derivative isogenic cell lines by knocking out the Tsc1 or Tsc2 gene by CRISPR/Cas9, and the derivative lines were designated T1G and T2J, respectively (Bissler et al., 2019 (link)). The Tsc1 gene was knocked out by targeting exon 4 (Bissler et al., 2019 (link)). Cystic kidney-derived epithelial cells, i.e., M1 cells, were also purchased from ATCC^® (CRL-2038^TM, Manassas, VA, United States). mIMCD3, T1G, and T2J cells were maintained in DMEM/F12 with 10% FBS, whereas M1 cells were maintained with DMEM/F12 with 5% FBS and supplemented with 5 μM dexamethasone. Cultures were maintained at 37°C in a humidified 95% air and 5% CO₂ atmosphere.