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. 2016 Feb 5;6:20460. doi: 10.1038/srep20460

Figure 2. KCTD12 suppresses the stemness of CRC cells.

Figure 2

(A) KCTD12 protein was analyzed by Western blotting in the indicated CRC cell lines. (B) The indicated stable cell lines with silencing or overexpression of KCTD12 were analyzed by Western blotting. α-tubulin or HSP70 was used as the loading control. (C-E) CD44, CD133 and CD29 levels were analyzed by Western blotting, qRT-PCR and flow cytometry, in the indicated stable cell lines. Red lines indicating the mean intensity of fluorescence of CD44+ or CD133+ were quantified by Flow-J software in the flow cytometry analysis. The mean intensity of fluorescence of CD44+ or CD133+ was calculated in triplicates. (F,G) Images and quantification of the number and size of spheres formed from the indicated stable cell lines in the absence of serum for 7 days. Original magnification in F, 40×(upper), 400×(lower). Original magnification in G, 40×. Scale bars, 100 μm. The results are presented as the means ± SD, and all data are representative of three independent experiments. *P < 0.05, **P < 0.01.