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. 2020 Nov 25;48(22):12711–12726. doi: 10.1093/nar/gkaa1090

Figure 5.

Figure 5.

USP36 regulates K29-linked polyubiquitination of PrimPol. (A) Knock-down of USP36 promotes K29-linked polyubiquitination of PrimPol. HEK293T cells stably expressing control (Ctrl) or USP36 shRNAs were transiently transfected with HA-tagged Ub-K29 and Myc-tagged PrimPol. After 48 h, cells were treated with MG132 (50 μM) for 3 h before collecting. Cell lysates were subjected to immunoprecipitation with anti-Myc agarose beads, and then blotted with the indicated antibodies. (B) USP36 stably knock-down HEK293T cells were transiently transfected with indicated constructs. After 48 h, cells were treated with MG132 (50 μM) for 3 h before collecting. Cell lysates were subjected to immunoprecipitation with anti-Myc agarose beads, and then blotted with the indicated antibodies. (C) Ctrl or USP36 stably knock-down HEK293T cells were transiently transfected with indicated HA-K29 lysine-specific mutant constructs and Myc-tagged PrimPol. After 48 h, cells were treated with MG132 (50 μM) for 3 h before collecting. Cell lysates were subjected to immunoprecipitation with anti-Myc agarose beads, and then blotted with the indicated antibodies.