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. 2023 Nov 15;7(2):e202302247. doi: 10.26508/lsa.202302247

Figure 7. DOCK1 interacts with DUSP4 and impairs its stability.

Figure 7.

(A) DUSP4 protein levels in DOCK1-overexpressing HTR-8 cells. (B) Interaction of DOCK1 and DUSP4 detected by CO-IP in HEK293T cells transfected with ectopic Flag-DOCK1 and Myc-DUSP4. (C) Interaction of DOCK1 and DUSP4 in the cytoplasm observed by immunofluorescence staining with anti-DOCK1 (red) and anti-DUSP4 (green) in HTR-8 cells. Scale bar: 25 μm. (D) DOCK1 knockout HTR-8 cells were treated with 50 μg/ml cycloheximide (CHX) for 0, 1.5, 3, and 6 h, and DOCK1 and DUSP4 proteins were assayed by Western blot. (E) HTR-8 cells overexpressing DOCK1 were treated with 50 μg/ml CHX for 0, 1.5, 3, and 6 h, and Western blot was used to detect DOCK1 and DUSP4 proteins. (F) HTR-8 cells were treated with 50 μg/ml CHX alone or 50 μg/ml CHX and 20 μM MG132 for 6 h. (G) HTR-8 cells were treated with 50 μg/ml CHX alone or 50 μg/ml CHX and 10 μM CQ or 50 μg/ml CHX plus 1 mM 3-MA for 6 h. (H) DOCK1-overexpression and control HTR-8 cells were treated with 20 μM MG132 or left untreated for 6 h.