Puromycin treatment of cells produces puromycinylated CNX-associated products and induces RESET of YFP-PrP* by a mechanism that is dependent on new protein synthesis. (A) Western blots depicting column purifications of endogenous CNX from lysates of NRK cells treated with 50 μg/ml cycloheximide and 2.50 µg/ml of puromycin (“CHX+Puro”), untreated (“U”), or treated with puromycin with 2.50 µg/ml of puromycin alone (“Puro”). Blots were probed with anti-CNX (αCNX) antibody to detect endogenous CNX and anti-puromycin antibody to detect puromycinylated proteins products that co-eluted with CNX. Input “I” and eluate “E.” (B) Time-lapse imaging of representative YFP-PrP* and Cerulean (CER)-GalT NRK cells after treatment with 2.50 µg/ml of puromycin alone (“+Puromycin”) or together with 50 μg/ml cycloheximide (“+Puromycin+CHX”), CER-GalT is a Golgi marker. Source data are available for this figure: SourceData F8.