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. 2021 Apr 7;7(15):eabe2261. doi: 10.1126/sciadv.abe2261

Fig. 4. Deubiquitinase USP7 interacts with methylated FOXA1 to remove FOXA1 protein ubiquitination.

Fig. 4

(A) LNCaP cells were infected with either shCtrl or two independent shUSP7 lentiviruses (shUSP7-1 and shUSP7-2) and subjected to WB. (B) 293T cells were transfected with FLAG-USP7, either alone or together with FOXA1. Co-IP was performed with a FOXA1 antibody. (C) 293T cells were transfected with the indicated plasmids and treated with 20 μM MG132 for 16 hours before co-IP by anti-FOXA1 and WB by anti-HA (Ub). (D) LNCaP cells stably expressing the indicated proteins were transfected with HA-Ub for 72 hours, treated with 20 μM MG132 for 16 hours, and then subjected to co-IP. (E) 293T cells were transfected with control, FLAG-FOXA1, FLAG-K295A, or FLAG-K295R. Co-IP was performed with anti-FLAG M2 beads. (F) 293T cells were transfected with FLAG-FOXA1 along with different HA-EZH2 constructs. Co-IP was performed using anti-FLAG M2 beads. (G) Co-IP using LNCaP nuclear lysates was performed with IgG, FOXA1, or K295me1 antibody.