Fig 2: High levels of HDGF correlate with poor survival of patients in HCCs with inactive Wnt signaling. A, B HDGF expression values and survival data of the TCGA-LIHC cohort were retrieved from OncoLnc [32] and stratified into Wnt-inactive (A) and Wnt-active (B) HCCs according to Sanchez-Vega et al. [33]. Patients were grouped into low (lower median) or high (upper median) expressions of HDGF. Kaplan–Meier with log-rank test

Fig 3: HDGF knockdown exerts distinct tumor-suppressive effects in Wnt-inactive HCC cells. A HLE and HepG2 cells were transfected with 10 nM siRNA against HDGF. HDGF expression was analyzed 48 h after siRNA transfection by qRT-PCR. Results were normalized to si-control. ***p < 0.001; one-way ANOVA with Dunnett’s multiple comparisons test. B HDGF and β-catenin protein expression was determined 48 h after siRNA transfection by western blotting with cofilin as loading control. Densitometric analysis of western blot assays is shown in Additional file 1: Fig. S9C. C HLE and HepG2 cells were transfected with 10 nM siRNA. Cell viability was analyzed by WST-1 assay and normalized to si-control (dotted line). Apoptosis was analyzed by Caspase 3/7 assay and normalized to cell viability and si-control (dotted line). *p < 0.05; two-way ANOVA with Dunnett’s multiple comparisons test. D Migration capacity of HLE cells was analyzed by transwell assay. **p < 0.01; one-way ANOVA with Dunnett’s multiple comparisons test; Scale bar = 200.00 µm. E HLE and HepG2 cells were transfected with 10 nM siRNA against HDGF. 48 h after transfection, protein expression of p-ERK1/2 and ERK1/2 was analyzed by western blotting with α-actinin as loading control. Gels were processed in parallel. Densitometric analysis of western blot assays is shown in Additional file 1: Fig. S9D

Fig 4: HDGF is a direct target gene of miR-129-5p. A Represented are predicted binding sites of miR-129-5p in the 3′UTR of HDGF. Binding sites were predicted by TargetScan [29] and IntaRNA [30]. The seed region of miR-129-5p is highlighted in gray. B Firefly luciferase activity was measured and normalized to renilla luciferase activity. ***p < 0.001; one-way ANOVA with Dunnett’s multiple comparisons test. C HDGF expression was analyzed 24 h, 48 h, and 72 h after transfection of HCC cell lines with 50 nM miR-129-5p mimics by qRT-PCR using the ΔΔCT method. *p < 0.05, **p < 0.01, ***p < 0.001; two-tailed Student’s t test. D HDGF protein expression was analyzed 48 h after miR-129-5p transfection by western blotting with cofilin as loading control. Densitometric analysis of western blot assays is shown in Additional file 1: Fig. S9B