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. 2018 Apr 30;128(6):2281–2296. doi: 10.1172/JCI93910

Figure 7. Requirement of posttranslational modifications for REV-ERBα degradation.

Figure 7

(A–C) Ubiquitinated REV-ERBα protein in HEK293T cells transfected with HA–Rev-Erbα, His-Ub, and SENP-1 plasmids and treated with GSK1362 at 10 μM, roscovitine at 25 μM, and TNF-α or IL-1β at 5 ng/ml for 4 hours in the presence of MG132 at 5 μM. (D–G) SUMO-2 ligation to REV-ERBα protein in HEK293T cells transfected with HA–Rev-Erbα, His-SUMO2, Ubc9, and SENP-1 plasmids and treated with GSK1362 at 10 μM and kinase inhibitors and IL-1β or TNF-α at 5 ng/ml for 4 hours in the presence of MG132 at 5 μM. (H) Coimmunoprecipitation of HDAC3 and REV-ERBα protein in HEK293T cells transfected with HA–Rev-Erbα and SENP-1 plasmids and treated with GSK1362 at 10 μM and IL-1β at 5 ng/ml for 4 hours. All blots are representative of at least n = 3.