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. 2019 Aug 9;10(8):603. doi: 10.1038/s41419-019-1838-0

Fig. 2.

Fig. 2

SOX2 knockdown inhibits CSCs self-renewal and maintenance in HNSCC. a Efficient SOX2 knockdown mediated by siRNA transfection in Cal27 and Fadu cells was confirmed by western blot assay. b, c Expression of multiple CSCs biomarkers including CD44, CD133, OCT4 and ALDH1A1 was measured by immunofluorescence staining (b, Scale bar: 50μm) and western blot (c). d, e Impaired tumorsphere formation was observed in cells transfected with two independent siRNAs targeting SOX2. Scale bar: 100 μm. Data were presented as Mean ± SD from three independent experiments, *P < 0.05, **P < 0.01, ANOVA analyses. f The protein abundance of TAZ and SOX2 was measured by western blot in both CSCs (CD44+CD133+) and non-CSCs (CD44CD133) subpopulations isolated from Cal27. g Tumorsphere was induced to differentiation into monolayer by serum from day 1 to 7. Representative images were shown. Scale bar: 100 μm. h The protein abundance of TAZ and SOX2 was measured by western blot in tumorsphere and serum-induced differentiated monolayer cells. i Tumor initiation was determined when cells transfected with SOX2 shRNAs were limitedly diluted and inoculated subcutaneously into NOD/SCID mice. Tumor was monitored and counted until 6 weeks after xenograft. Tumor-initiating frequencies were calculated via ELDA software (http://bioinf.wehi.edu.au/software/elda/). Data were presented as Mean ± SD from three independent experiments, *P < 0.05, **P < 0.01, ANOVA analyses