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. 2015 Sep 10;6(31):31927–31943. doi: 10.18632/oncotarget.5578

Figure 4. Stat3 is critical for HCC initiation and is involved in Icaritin-reduced hepatosphere formation.

Figure 4

(A). PLC/PRF/5 and Huh7 cells were treated with DMSO control or 150 μM of S3I-201 for 48 h. Western blots were performed with the indicated antibodies. (B). PLC/PRF/5 and Huh7 cells were treated with DMSO control or 150 μM of S3I-201 for 48 h. The living cells were sorted with FACS and used for the hepatosphere formation assay in the absence of S3I-201. (C). PLC/PRF/5 cells were transfected with control siRNA or the Stat3 siRNA for 48 h. Total cell lysates were prepared and analyzed with western blots using the indicated antibodies. (D). PLC/PRF/5 cells were transiently transfected with the control siRNA or the Stat3 siRNA for 48 h. The transfected cells were then used for hepatosphere formation assay in the presence of the indicated concentrations of Icaritin. Data are expressed as percent of the control. (E). PLC/PRF/5 cells were transiently transfected with the vector control or the Stat3-C expression plasmid for 48 h. Cells were harvested for western blots assay and hepatosphere formation assay. Data are expressed as percent of the vector control. Columns, mean (n = 3); bars, SEM. *, P < 0.05; **, P< 0.01; ***, P<0.001; ICT, Icaritin.