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. 2022 Jul 11;12:899987. doi: 10.3389/fonc.2022.899987

Figure 4.

Figure 4

USP13 inhibits polyubiquitylation-dependent FASN degradation. (A) H1048 (upper) and H69 (lower) cells expressing two different USP13 shRNAs were treated with MG132 (50 μM) for 8 hours. Immunoblot analyses were performed with the indicated antibodies. (B) H1048 cells with or without USP13 depletion were treated with CHX (100 µg/ml) for the indicated periods of time. FASN expression levels were analyzed with an anti-FASN antibody (upper) and quantification of FASN levels relative to β-actin expression levels (lower) were performed. (C) H1048 cells with or without WT USP13 or catalytically inactive USP13 (C345A) mutant overexpression were treated with CHX (100 µg/ml) for the indicated periods of time. Immunoblot analyses were performed with the indicated antibodies (left). Quantification of FASN expression levels relative to β-actin expression levels is shown (right). (D) H1048 cells expressing two different USP13 shRNAs or a control shRNA were treated with MG132 (50 μM) for 8 hours. Immunoprecipitation and immunoblot analyses were performed with the indicated antibodies. (E) H1048 cells with or without WT USP13 or catalytically inactive USP13 (C345A) mutant overexpression were treated with MG132 (50 μM) for 8 hours. Immunoprecipitation and immunoblot analyses were performed with the indicated antibodies.