Research Paper Volume 13, Issue 15 pp 19317—19338

The deubiquitinase OTUD3 stabilizes ACTN4 to drive growth and metastasis of hepatocellular carcinoma

OTUD3 is significantly upregulated in human HCC tissues. (A) Representative images for OTUD3 IHC staining in HCC tissues and corresponding normal tissues (N=115; scale bar: 50μm, 100μm). (B) Diagram of OTUD3 staining score in IHC staining. (C) qRT-PCR analysis of OTUD3 mRNA level in 50 cases of HCC tissues and corresponding normal tissues. Left, a log 2 (T/N) value> 0 indicates that OTUD3 expression is overexpressed in the HCC samples; right, a log 2 (T/N) valueD) Determination and quantification of OTUD3 protein levels in HCC tissues and paired non-tumour tissues by western blot (N=50). (E) Representative images of western blot. (F) Kaplan–Meier analysis of the correlation between the OTUD3 level and overall survival of HCC patients with high and low OTUD3 expression in the IHC staining. (G) Kaplan–Meier analysis of OTUD3 expression in 364 liver cancer tissues using the Kaplan–Meier plotter website (<a href="https://www.kmplot.com" target="

Figure 1. OTUD3 is significantly upregulated in human HCC tissues. (A) Representative images for OTUD3 IHC staining in HCC tissues and corresponding normal tissues (N=115; scale bar: 50μm, 100μm). (B) Diagram of OTUD3 staining score in IHC staining. (C) qRT-PCR analysis of OTUD3 mRNA level in 50 cases of HCC tissues and corresponding normal tissues. Left, a log 2 (T/N) value> 0 indicates that OTUD3 expression is overexpressed in the HCC samples; right, a log 2 (T/N) value< 0 indicates that OTUD3 expression is downregulated in the HCC samples. The OTUD3 mRNA levels are normalized to the GAPDH mRNA levels. (D) Determination and quantification of OTUD3 protein levels in HCC tissues and paired non-tumour tissues by western blot (N=50). (E) Representative images of western blot. (F) Kaplan–Meier analysis of the correlation between the OTUD3 level and overall survival of HCC patients with high and low OTUD3 expression in the IHC staining. (G) Kaplan–Meier analysis of OTUD3 expression in 364 liver cancer tissues using the Kaplan–Meier plotter website (https://www.kmplot.com). Data were mean ± S.D. of three independent determinations. Student’s t-test was used for P value assessment. *P< 0.05, **P<0.01.