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. 2021 Dec 10;118(50):e2107993118. doi: 10.1073/pnas.2107993118

Fig. 3.

Fig. 3.

K63, K27-linked ubiquitin chain reprimation induces autophagic degradation of the Cys substrates. (A) Denaturation IP of RGS4MYC and HA-Ub expressing HEK293T cells treated with a combination of BAF (200 nM), tBHP (250 μΜ), and MG132 (10 μΜ) for 6 h. (B and C) Same as A but with HA-Ub, and (B) C and RC-RGS4FLAG expressing ATE1−/− MEF cells or (C) RC-RGS4FLAG and treated with MG132 or BAF with tBHP. (D) Same as C but with tBHP and BAF only treatment. (E) Immunocytochemistry in SH-SY5Y cells expressing K63 only and K27 only HA-Ub treated with BAF, tBHP, or both (same as A). (F–I) Denaturation IP of HEK293T cells expressing RGS4MYC (F, H, I) or RGS16MYC (G) with a combination of K63 only (F and G), K27 only (H), or K48 only (I) HA-Ub mutant treated with tBHP, BAF, or both (same as A).