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. Author manuscript; available in PMC: 2016 Jun 1.
Published in final edited form as: Stem Cells. 2015 Jun;33(6):1705–1718. doi: 10.1002/stem.1993

Figure 2. YAP1 silencing abrogates the self-renewal ability of CSCs.

Figure 2

(A) Sphere formation assay with SP cells from A549 and H1650 transfected with two different siRNA against YAP1 show smaller spheres as compared to a non-target control siRNA (B) Average number of spheres generated from 1000 SP cells reveal fewer number of spheres in YAP1 siRNAs treated cells as compared to the control siRNA in both cell lines. (C) Quantitation of serial sphere formation assay show fewer spheres in SP cells isolated from A549 and H1650 cells transfected with YAP1 siRNAs as compare to control siRNA treatment. Bright field images of the spheres are presented here. (D) SP cells isolated from YAP1 siRNAs treated H1650 cells show abrogation of angiogenic tubule-like structure formation when grown on Matrigel in endothelial growth medium. (E) Loss of CD31 expression on angiogenic tubule-like structures in SP cells from YAP1 siRNA treated H1650 as visualized by immunofluorescence. (F) Real time PCR analysis of YAP1 siRNA treated H1650 cells show a decrease in CD31 mRNA expression as compared to control siRNA treated cells. (G–H) Real time PCR analysis of YAP1 siRNAs transfected A549 (G) and H1650 (H) cells respectively for YAP1, Sox2, Oct4, Nanog and ABCG2 genes show decrease in Sox2, Oct4, Nanog expression. The above data is expressed as mean ± SD of three independent experiments. * represents p < 0.05, ** represents p < 0.01.