Fig. 3: Transcription factor Ets-1 expression is induced in soraR cells.

A Total RNA extracted from Hep3B-naive and soraR cells was subjected to cDNA synthesis and analyzed with human transcription factors PCR Array (PAHS-075Z). Figure in A shows the heat map with the upregulated genes marked in red and the downregulated genes marked in green. B shows the scatter plot of expression levels of 84 genes. Red dots indicate genes upregulated and the green dots indicate the genes downregulated. The central line indicates genes that were unchanged (black dots) with dotted lines indicating the selected twofold regulation cut-of. C Changes in the expression of ETS-1 gene in Huh7 and Hep3B (naive and soraR) cells were analyzed by qPCR. The data represent the mean ± S.D. of 3 independent PCR reactions, and the experiment was repeated three times. D Equal amounts of total protein from 3 sets of naive (N) and soraR (SR) HCC cells were analyzed by western blots with antibodies against Ets-1 and GAPDH. E Immunocytochemistry showing differences in expression of Ets-1 in naive and soraR cells. The images were captured using the NIS-Elements imaging software in Nikon Eclipse Ti Microscope. Scale bar, 20 µm. F Representative images of immunohistochemistry of Ets-1 using liver cancer tissue microarray (US Biomax, # T031b). Scale bar, 60 µm. The Ets-1 staining intensities in the TMA were scored and plotted to the right. Statistical significance was determined by Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: p > 0.05 not significant. HCC-CC hepatocholangiocarcinoma.