Fig. 4. Effect of the combination treatment of NVP-BKM120 and Prima-1Met on stem-like traits of thyroid cancer cells and the expression of thyroid-specific differentiation markers. (a) THJ-29T and THJ-21T cells were grown in 3D, treated with drugs as indicated for 14 days, and quantitated for structural integrity. (b) Sphere formation efficiency of the cells in (a) was calculated. (c) Protein level of CD44, CD133, OCT4, ABCG2 and Nanog were determined in THJ-29T and THJ-21T cells treated with BKM120, Prima-1Met or the two in combination. (d) mRNA level of NIS and Tg were determined in THJ-29T and THJ-21T cells treated with BKM120, Prima-1Met or the two in combination. (e) The expression level of NIS in THJ-29T and THJ-21T cells treated with BKM120, Prima-1Met or the two in combination were analyzed by Western blot.