Pbabe nf2

A 1.5-kb promoter region of the murine Nf2 promoter containing the Sox2-binding site was cloned into the HindIII site of the pGL3-Basic Vector (Promega, WI, USA). The Sox2-binding site in the Nf2 promoter was mutated using the Quick Change II Site-Directed Mutagenesis Kit from Agilent technologies, Santa Clara, CA. A 750-bp fragment of the 3′UTR of the murine WWC1 gene was cloned into the XbaI site of the pTK-Luciferase plasmid. The 8X-GTIIC TEAD luciferase reporter was a kind gift from Dr Stefano Piccolo. Nf2 shRNAs (mouse), Sox2 shRNAs (human) and YAP shRNAs (mouse) lentivirus were purchased from Sigma, St Louis, MO. The Nf2 lentivirus was a kind gift from Dr Filippo Giancotti. The FUCRW-Sox2 and FUCRW-YAP lentiviruses have been previously described^{29 (link)}. Other retrovirus such as Myc-tagged WT YAP (Addgene # 33091), 5SA YAP (Addgene # 33093)and 94SA YAP (Addgene # 33094) all in the pQCXIH vector from the laboratory of Dr Kung-Lian Guan, and pBabe-Nf2 (Addgene # 14116) from the laboratory of Dr Tyler Jacks, were obtained from Addgene, Cambridge, MA. All lentiviruses used in this study were produced in 293T cells using pVSV-G, pLP1 and pLP2 as helper plasmids. All retroviruses were produced in 293-GP cells and pseudotyped with VSV-G.

pBabe-NF2 was obtained from Addgene. Anti-Fasn siRNA (M-040091-01-0005), anti-Acaca siRNA (M-063938-01-0005), anti-Mtor (M-065427-00-0005), anti-Rptor (M-058754-01-0005), anti-Rictor (M-064598-01-0005) and non-silencing (D-001206-13-05) siRNA were purchased from Dharmacon. Individual siRNAs against Mtor (SASI_Mm01-00164496 and -00164492), Rictor (-00137732 and -00137730), Rptor (-00055298 and -00334580), Fasn (-00177858 and -00177854), Acaca (-0011590 and -00115905), and Mlycd (-00028572 and -00028576) were purchased from Sigma-Aldrich. Anti-Merlin antibodies were purchased from Abcam (#ab88957). Lipid synthesis and metabolism antibody kit (includes anti-Fasn, -phospho ACC, -ACC, -Lipin1, -ACLY, -phospho ACLY, -ACSL1, and -ACECS1 antibodies), and anti-Casp3 antibodies were purchased from Cell Signaling Technology. Anti-SREBP1 antibodies were purchased from Santa Cruz Biotechnology. Anti-GAPDH antibodies were purchased from EMD-Millipore.
Cerulenin, C75, luteolin, 5-(tetradecyloxy)-2-furoic acid (TOFA) and 5-iodotubercidin were purchased from Enzo Life Sciences. GSK2194069, dimethylsulfoxyde (DMSO), staurosporin, sodium palmitate, 70% perchloric acid, ammonium formate, acetonitrile, acetyl-coenzyme A lithium salt, malonyl coenzyme A lithium salt, propionyl-coenzyme A lithium salt, and poly-L-lysine were purchased from Sigma-Aldrich.