For immunoblotting, antibody against ubiquitin (Ub), AMPK‐α1, p‐AMPK (Thr172), Akt, p‐Akt (Thr308), p‐Akt (Ser473), ERK1/2, LC3‐I/II, or p‐CREB (Ser133) was purchased from Cell Signaling Technology (Danvers, MA, USA). Anti‐MAGE‐A3, anti‐ASS1, anti‐p‐ERK, anti‐AMPK‐β, anti‐AMPK‐r, anti‐TRIM28, and anti‐CREB antibodies were separately, obtained from Abgent (San Diego, CA, USA), BD Biosciences (San Jose, CA, USA), Polaris, Sigma‐Aldrich, and GeneTex (Irvine, CA, USA). Anti‐RNF44 antibody was purchased from Abcam (Cambridge, MA, USA). All secondary antibodies conjugated with HRP were purchased from Promega (Madison, WI, USA). The immunoblots were developed using ultrasensitive enhanced chemiluminescent substrate and visualized by ChemiDoc MP System (Bio‐Rad, Hercules, CA, USA). The anti‐CREB antibody used for chromatin immunoprecipitation (ChIP) was purchased from Cell Signaling Technology.
For CAT‐1 and CAT‐2 detection, melanoma cells were incubated with primary antibody (CAT‐1, Novus, Littleton, CO, USA, 1 : 20; CAT‐2, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 20) for 20 min and then incubated with second antibody conjugated with Alexa Fluor® 555 (Thermo Fisher Scientific) for 20 min at room temperature. The samples were analyzed by FACS (BD Accuri™ C6).
For CAT‐1 and CAT‐2 detection, melanoma cells were incubated with primary antibody (CAT‐1, Novus, Littleton, CO, USA, 1 : 20; CAT‐2, Santa Cruz Biotechnology, Santa Cruz, CA, USA, 1 : 20) for 20 min and then incubated with second antibody conjugated with Alexa Fluor® 555 (Thermo Fisher Scientific) for 20 min at room temperature. The samples were analyzed by FACS (BD Accuri™ C6).