Fig. 4
Effect of combinatorial treatment with SLC-0111 and Cis-Pt on tumorsphere formation and stemness. FaDu (A) and SCC-011 (B) cells were seeded in Ultra-Low attachment multiwell-plates and grown in serum free DMEM supplemented with B27, bFGF (20 ng/mL) and EGF (10 ng/mL) in hypoxic conditions for 7 days. HNSCC cells were treated with Cis-Pt (1 µM), SLC-0111 (100 µM) and combination of the two drugs for 72 h. Spheroid formation was analyzed under a phase-contrast microscopy and size and number of formed spheroids was calculated using ImageJ. All the data are expressed as percentage considering the untreated control cells as 100%. Bars depict mean ± SD of three independent experiments (*** p < 0.0001; ** p < 0.001; *p < 0.01). C Western blot analysis of expression levels of stemness markers Nanog and Sox-2 in FaDu and SCC-011 grown under hypoxia (1% O2) and treated with Cis-Pt (1 µM), SLC-0111 (100 µM) and combination of the two drugs for 72 h. Tubulin was used as loading control. Representative data from one of three experiments are shown