hTERT RPE1 (retinal pigment epithelial) and HEK293T cells were purchased from ATCC. hTERT RPE1 COG4 KO cells were described previously.29 (link) Cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing Nutrient mixture F-12 (DMEM/F12, Corning 10–092-CV) supplemented with 10% fetal bovine serum (Atlas Biologicals, CF-0500-A). Cells were incubated in a 37°C incubator with 5% CO2 and 90% humidity.
For rapid COG4 degradation, a stock solution of 0.5 M Indole-3-acetic acid sodium salt (auxin, IAA, Sigma # I5148) was prepared in water and stored in a frozen aliquot. Time course treatment of cells was performed with 500 μM IAA for 0.5, 1, 2, 24, and 48 h at 37°C. The cells without auxin treatment were considered as untreated control.
For rapid COG4 degradation, a stock solution of 0.5 M Indole-3-acetic acid sodium salt (auxin, IAA, Sigma # I5148) was prepared in water and stored in a frozen aliquot. Time course treatment of cells was performed with 500 μM IAA for 0.5, 1, 2, 24, and 48 h at 37°C. The cells without auxin treatment were considered as untreated control.
