Fig 2: Upregulations of TIPRL, LC3 and CD133 in HCCs. Human tissues were stained with the indicated antibodies followed by confocal observation. (a) The levels of TIPRL, LC3 and CD133 were determined using the ZEN program (Supplementary Tables 1 and 2). p-values (*P < 0.05; **P < 0.01) were determined by a paired t-test, and % differences are shown. (n) is the number of samples. (b) The representative images were selected from normal and HCCs groups, respectively. DAPI was used for nucleus staining, and scar bars, 20 (inserted) and 100 µm.

Fig 3: Diagnostic efficacies of TIPRL, LC3, CD133 and the TIPRL/LC3/CD133 models in HCCs and early liver cancers. Diagnostic efficacies of TIPRL, LC3, CD133 and the combined models in HCCs (a,b) and grade1 liver cancers (c,d) were determined using ROC analysis. AUC, area under the curve.

Fig 4: Significant correlations between levels of TIPRL, LC3 and CD133 in liver cancers. (a–c) The statistically significant associations between levels of TIPRL, LC3 and CD133 in each grade of liver cancers were determined by the Spearman correlation. Each dot indicates a single patient. (d) The representative images were selected from each grade of human liver cancer tissues that were stained with indicated antibodies. DAPI was used for nucleus staining, and scar bar, 100 µm. (e) Quantitative analyses of TIPRL, LC3 and CD133 were conducted according to each grade of liver cancers. (n), the number of samples is indicated.

Fig 5: Confirmation of reproducibility of TIPRL, LC3 and CD133 as HCC biomarkers. (a) Using a different IHC cohort (Supplementary Tables 4 and 5), the levels of TIPRL, LC3, CD46 and CD133 were quantified using the ZEN program. Statistical differences (*P < 0.05; ***P < 0.0001) were calculated by a paired t-test, and % differences are shown. (n), the number of samples is indicated. (b–f) The correlations between levels of TIPRL, LC3, CD46 and CD133 were determined by the Spearman correlation equation. Single dots indicate an individual sample. (g) Diagnostic efficacies of TIPRL, LC3, CD133 and CD46 were determined by the ROC analysis. AUC, area under the curve. (h) Our study shows that the combination of TIPRL contributing to drug resistance with the cancer stem marker CD133 and autophagy marker LC3 produces novel HCC biomarkers. These biomarkers show more significant diagnostic efficacies than the CD46 currently being used as a liver cancer marker.