Fig 2: DAAM1 is coprecipitation and colocalized with YHWAZ.A, B The expression of exogenous proteins in MCF-7 and MDA-MB-231 BrCa cells transfected with the indicated constructs was examined by western blotting. C, D Endogenous FLAG-DAAM1 and GFP-YWHAZ coimmunoprecipitated with each other with the anti-FLAG or anti-GFP antibody. The whole-cell lysates were analyzed by immunoblotting with the indicated antibodies. E Subcellular localization of DAAM1 and YWHAZ was analyzed by immunofluorescence staining followed by capturing using a laser scanning confocal microscope (Olympus FV3000). Magnification: ×600, Bar = 40 μm.

Fig 3: YWHAZ impairs UBR5-suppressed liver cancer cell proliferation (A) UBR5 and YWHAZ protein levels were detected, respectively, by western blot in Huh7 and Hep3B cells as a quality control after UBR5 knockdown (siRNA-1, 2) with siRNA in vitro. MOCK and NC, respectively, served as blank and negative controls. Quantification of results (right). (B) Western blot analysis of YWHAZ expression of YWHAZ-OE in siUBR5 (siRNA-2) transfected HCC cells or controls. Quantification of results (right) (C,D) cell growth at different time points (every 1 day interval) was analyzed using an MTT assay in Huh7 and Hep3B cells. YWHAZ-OE, YWHAZ Overexpression. *p < 0.05.

Fig 4: The level of YWHAZ expression in liver cancer and normal liver tissues. (A) YWHAZ is more abundant in liver cancer than in normal liver tissues. ***p < 0.001. (B) The association between YWHAZ expression and overall survival in patients with liver cancer. Kaplan-Meier survival analysis of UBR5 expression in gastric cancer patients. High YWHAZ expression was attributed to shorter survival time, while low YWHAZ expression was correlated to longer survival time (p = 0.00012).

Fig 5: UBR5 co-expression genes in liver cancer. (A,B) Heat map revealed 24 important genes that were favorably and negatively associated with UBR5 respectively. (C,D) The pearson test was performed to identify genes in liver cancer that were differentially expressed in response to UBR5. (E) PPI network revealed that UBR5 had a close association with YWHAZ. PPI, protein–protein interaction.