Fig. 2

Silencing TM4SF1 could inhibit the cancer stemness in HCC in vitro. A Stemness index of TM4SF1 in 371 HCC samples from the TCGA database was calculated using the OCLR algorithm, and the degree of stemness correlation of TM4SF1 was evaluated. B The connection between TM4SF1 and chemicals associated to cancer stem cells was examined using the GEPIA database. C RNA was extracted from Hep3B tumor spheres and detected via qPCR. The expression of TM4SF1 in tumor balls abnormally increased. D qPCR was used to verify the knockout efficiency of TM4SF1 in Hep3B and LM3 after transient transfection. E WB was used to confirm the knockout efficiency of TM4SF1 in Hep3B and LM3 after transient transfection. F qPCR was used to verify the knockout efficiency of TM4SF1 in Hep3B and LM3 after lentivirus infection. G WB was used to verify the knockout efficiency of TM4SF1 in Hep3B and LM3 after lentivirus infection. H The outcomes of qPCR analysis revealed that TM4SF1 silencing reduced the expression of markers linked to cancer stem cells. I The outcomes of WB analysis revealed that TM4SF1 silencing reduced the expression of markers linked to cancer stem cells. J Results of the experiment for the detection of side-population cells showed that after silencing TM4SF1, the proportion of stem cells in hepatocellular carcinoma cells decreased significantly. K The ratio of CD44 positive and CC133 positive cells in TM4SF1 silenced LM3 and control group was determined by fluorescence-activated cell sorting. L Experiment for tumor sphere formation showed that the number and diameter of tumor spheres decreased after TM4SF1 was silenced. Scale bars, 50 μm. M Comparison of the number of tumor spheres and the proportion of tumor spheres with different diameters. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001