Fig. 4: Nobiletin prevented the interactions of AR-V7 with USP14 and USP22.
From: Selective degradation of AR-V7 to overcome castration resistance of prostate cancer
A Co-IP assays using AR-V7 antibody and immunoblot to indicated deubiquitinases in 22Rv1 cells. B Immunofluorescence assays using HA and USP14 antibodies in 22Rv1 cells transfected with HA-AR-V7 plasmids for 48 h. Scale bars represent 10 μm in shown images. C Immunofluorescence assays using HA and His antibodies in 22Rv1 cells co-transfected with His-USP22 and HA-AR-V7 plasmids for 48 h. Scale bars represent 10 μm in shown images. D Western blot analysis of AR, AR-V7 USP14, and USP22 protein level in cytoplasmic and nuclear of 22Rv1 cells treated with nobiletin for 24 h. HSP90 and Lamin B1 were used as cytoplasmic and nuclear internal controls, respectively. E Co-IP assay was performed to determine the interaction of USP14/USP22 and AR/AR-V7 in 22Rv1 cells exposed to nobiletin for 12 h. F Co-IP assay was performed to determine the interaction of USP14/USP22 and AR/AR-V7 in 22Rv1 cells exposed to nobiletin (20 μM) for different lengths of time. G Co-IP assay was performed to determine the interaction of GRP78 and AR-V7 in 22Rv1 cells exposed to nobiletin for 12 h.
