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. 2016 May 26;291(29):15256–15266. doi: 10.1074/jbc.M116.722967

FIGURE 1.

FIGURE 1.

Identification of RNF146 and tankyrase as positive regulators of YAP signaling through pooled CRISPR screen. A, an illustration of FACS-based pooled CRISPR screen of YAP signaling pathway in HEK293T NF2 KO GTIIC-GFP cells. B, scatter plot depicting results of GTIIC-GFP pooled CRISPR screen. Y-axis indicates the RSA down value (the statistical significance of all gRNA targeting each gene being unusually distributed toward the low end of the distribution), and X-axis indicates the Q1 z-score. C, tankyrase inhibitors attenuate GTIIC-GFP YAP reporter in HEK293A cells. For FACS assay, data are representative from at least two independent experiments. D, tankyrase inhibitors decrease the expression of YAP target genes in HEK293A cells. For qPCR assay, error bars are S.D., n = 4. Data are representative from at least two independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; NS, not significant. E, tankyrase inhibitors attenuate GTIIC-GFP YAP reporter in HEK293T NF2 KO cells. F, tankyrase inhibitors decrease the expression of YAP target genes in HEK293T NF2 KO cells. G, depletion of TNKS1/2 decreases GTIIC-GFP in HEK293A cells. H, depletion of TNKS1/2 decreases the expression of YAP target genes in HEK293A cells. I, effect of tankyrase inhibitors on the level of pS127 YAP and total YAP in total cell lysates of HEK293A cells. J, tankyrase inhibitors decrease the level of YAP in nuclear extracts of HEK293A cells. K, tankyrase inhibitors decrease nuclear localization of YAP in HEK293A cells. L, tankyrase inhibitors decrease nuclear localization of YAP in HEK293T NF2 KO cells.