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. 2019 May 1;116(20):9877–9882. doi: 10.1073/pnas.1819400116

Fig. 3.

Fig. 3.

FASN-dependent regulation of YAP-TEAD activity. (A) Analysis of FASN and ACC expression. The expression of FASN and ACC proteins in MCF-10A at low and high cell densities was analyzed by Western blotting. H, high cell density; L, low cell density. (B) Quantitative analysis of FASN and ACACA mRNA expression. The expression of FASN and ACACA mRNAs in parental, and Lats1/2 or Nf2-depleted MCF-10A at low or high cell densities was analyzed using qPCR. (C) Biochemical effects of FASN depletion. Endogenous FASN was depleted by siRNA transfection in parental (10A), Lats1/2 (ΔLats), or Nf2 KO (ΔNf2) MCF-10A cells. Phosphorylation of YAP was monitored by phos-tag SDS/PAGE and Western blotting. (D) HOP-flash reporter. YAP-TEAD reporter assay was carried out in 293A cells transfected with control, Lats1/2, or Nf2 siRNAs in the presence/absence of FASN siRNA. (E) Quantitative analysis of YAP-TEAD target genes’ mRNA expression. The expressions of CTGF and Cyr61 mRNA in Lats1/2 and/or FASN-depleted MCF-10A cells were analyzed using qPCR.