Figure 4. YAP1 is required to sustain self-renewal and dedifferentiation of prostate cancer cells.

in vitro and in vivo. (A) LNCaP cells and C4-2 cell subpopulations were isolated using magnetic sorting with antibodies recognizing stem cell markers, CD133 and CD44. Protein levels of YAP1, CD133 and CD44 were analyzed by Western Blot in control and CD133 or CD44 high or low cells. (B) Quantitation of sphere formation capacity of C4-2, CD133^high cells transfected with sh-control or sh-YAP1 expression vectors. (C) Quantitation of sphere formation capacity of C4-2, CD133^low cells transfected with either control or YAP1-wt expression vectors. (D) mRNA expression of stem cell marker genes: CD44, CD133, SOX2, OCT4, Nanog, C-met and YAP1 by qPCR after siRNA mediated knockdown of YAP1 expression in C4-2 cells. (E) CD133^low cell, CD133^low-YAP1wt expressing cell and LNCaP cell were transplanted into nude mice subcutaneously and tumor growth was monitored daily. Growth curves are shown. P values were determined by paired t test. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001 (F) Quantitation of YAP1, Ki67 and CD133 protein expression in mice tumors using IHC. See also Supplementary Figure 2.