Fig. 4

TBC1D31 is a target of ZSCAN16 in HCC cells. A: The first 1500 downstream targets of ZSCAN16 downloaded from hTFtarget were intersected with DEGs (p. adj. < 0.01) in the GSE57957 and GSE46408 datasets obtained on Jvenn; B: UALCAN predicted the expression of TBC1D31 (WDR67) in HCC. C: Kaplan-Meier Plotter to analyze the prognostic significance of TBC1D31 (WDR67) in HCC. D: Correlation between ZSCAN16 and TBC1D31 in LIHC samples in the TCGA Tumor database and in liver samples in the GTEx database was analyzed on GEPIA. E: ChIP-seq database in the UCSC to analyze the binding of ZSCAN16 in the promoter region of TBC1D31. F: RT-qPCR to detect the transcription level of TBC1D31 in HCC tissues and paracancerous liver tissues. G: IHC to analyze the protein expression of TBC1D31 in HCC tissues. H: RT-qPCR to detect the expression of TBC1D31 in THLE-2 and HCC cell lines. I: WB detection of protein levels of TBC1D31 in THLE-2 and HCC cell lines. J: RT-qPCR detection of mRNA expression of TBC1D31 in HCC cell lines with ZSCAN16 knockdown. K: ChIP-qPCR detection of DNA fragments enriched by ZSCAN16 antibody. L: Dual-luciferase assay detected the relationship between ZSCAN16 and TBC1D31. All cellular experiments were independently repeated three times. The comparison between two groups was performed using the paired t-test (FG) or unpaired t-test (JL), and ANOVA analysis was used for comparisons among multiple groups (HIK). * p < 0.05 compared with adjacent tissue, THLE-2 cells, or the sh-Scramble group