Verteporfin

Cells were seeded at a density of 1 × 10⁴ cells/well in 96-well plates. When cells reached 60% confluence, drugs were added into the wells and incubated for 24 h or 48 h. Medium containing 10% Cell Counting Kit-8 (CCK-8) reagent (Dojindo, Japan) was then added into the cells and incubated for another 1.5 h at 37°C. The light absorbance was measured at 450 nm on the microplate reader (Bio-Rad, USA). Each group was performed in sextuplicate. Verteporfin, AZD-2858 were purchased from TargetMol, USA, and varlitinib was purchased from MCE, USA

Adriamycin was purchased from Sigma, USA, and was administrated in mT/mG mice at several different doses (15, 17, 19, 22 mg/kg body wt) by intravenous injection (one dose at 5 weeks of age). Verteporfin was purchased from TargetMol, USA and was administrated in mT/mG mice at 100 mg/kg body wt by intraperitoneal injection every other day for 1, 2, or 3 weeks (starting from the 1st day of the Adriamycin-induced nephropathy mouse model construction). Verteporfin dosages are consistent with a previous report [19 (link), 20 (link)]. The mice were then sacrificed.

LoVo and HCT116 cells were purchased from the Type Culture Collection of the Chinese Academy of Science (Shanghai, China). LoVo cells were cultured in RPMI 1640 cell culture (Gibco) containing 10% fetal bovine serum, and HCT116 cells were cultured in McCoy’s 5A (Gibco) containing 10% fetal bovine serum and cultured in a 5% CO₂ incubator at 37°C. The proteasome inhibitor MG132 (MedChemExpress, HY-13259) was used to treat HCT116 cells for 6 h, the YAP inhibitor verteporfin (TargetMol, T3112) was used to treat HCT116 cells for 16 h, and the YAP agonist PY-60 (TargetMol, T9566) was used to treat HCT116 cells for 24 h.