To generate SMURF2 knock-down, cells were infected with lentiviral particles containing one of the following shRNAs: shSMURF2#1 (targets SMURF2 at its coding sequence; Cat. #TRCN0000010792, Sigma, St. Louis, MO) and shSMURF2#2 (targets SMURF2 at 3′UTR; Cat. #TRCN0000003475, Sigma, St. Louis, MO). shRNA targeting luciferase (shLuc; Cat. #SHC007, Sigma, St. Louis, MO) served as a control12. Following infection, cells were selected with puromycin (1–2 µg/ml) for at least two weeks. SMURF2 knock-out cells were generated using an advanced CRISPR/Cas9 gene-editing tool, as previously described41. The efficiency of SMURF2 depletion was verified in immunoblots.

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