Supplementary Figures S1 - S10, Tables S1 - S3 from <i>NF2</i> Loss Promotes Oncogenic RAS-Induced Thyroid Cancers via YAP-Dependent Transactivation of RAS Proteins and Sensitizes Them to MEK Inhibition

Abstract

<p>Supplementary Figure S1. Loss of heterozygosity of Ch22q genes in poorly differentiated (PDTC) and anaplastic thyroid cancers (ATC). Supplementary Figure S2. Focal NF2-exon 4 homozygous deletion in KHM-5M anaplastic thyroid cancer cells. Supplementary Figure S3. FISH for NF2 in anaplastic thyroid cancers. Supplementary Figure S4. NF2/Merlin defects in thyroid cancer cell lines. Supplementary Figure S5. Merlin inhibits RAC1-PAK activity. Supplementary Figure S6. Merlin effects on EGFR signaling and growth of RAS mutant thyroid cancer cell lines. Supplementary Figure S7. PCR of genomic DNA from mouse thyroid tissues of the indicated genotypes with primers that distinguish wild-type from mutant Hras alleles. Supplementary Figure S8. TEAD1 is the most abundant TEAD isoform in thyroid cancer cells. Supplementary Figure S9. A) Left, Concentration-dependent growth inhibitory effects of the MEK inhibitor selumetinib (AZD6244) in RAS mutant thyroid cancer cell lines cells. Black, null/low NF2. Red, wt NF2. The IC50 of each cell line is shown in the boxed legend. Right, Western blots for merlin, pMEK and pERK in the cell lines studied. B) C643 cells expressing scrambled (pLKO.1) or shNF2 were treated with FRAX, AZD or their combination at the indicated concentrations (nM) in 1 percent of serum. Cells were counted at 6 days (*p<5 x E -3). Supplementary Figure S10. A) Tumor doubling time of the indicated mouse models of PDTC.B) Kaplan-Meier survival curves of TPO-Cre, TPO-Cre/FR-HrasG12V/NF2flox2, TPO-Cre/FR-HrasG12V/PTENflox2 and TPO-Cre/FRHrasG12V/p53flox2 mice. Left, HrasG12V heterozygotes. Right, HrasG12V homozygotes. (*p < 3 x E-2, **p < 1 x E -4). Supplementary Table S1. 22q loss is associated with RAS mutations in PTC, PDTC and ATC. Supplementary Table S2. Penetrance of thyroid cancer in HRASG12V/NF2-null mice. Supplementary Table S3. PCR primer sequences.</p>