Supplementary Material for:

A method of high-throughput functional evaluation of EGFR gene variants of unknown significance in cancer

Shinji Kohsaka,* Masaaki Nagano, Toshihide Ueno, Yoshiyuki Suehara, Takuo Hayashi, Naoko Shimada, Kazuhisa Takahashi, Kenji Suzuki, Kazuya Takamochi, Fumiyuki Takahashi, Hiroyuki Mano*

*Corresponding author. Email: kohsakas{at}m.u-tokyo.ac.jp (S.K.); hmano{at}m.u-tokyo.ac.jp (H.M.)

Published 15 November 2017, Sci. Transl. Med. 9, eaan6566 (2017)
DOI: 10.1126/scitranslmed.aan6566

This PDF file includes:

• Fig. S1. Quantification of focus-formation assay.
• Fig. S2. Temporal changes in the proportion of 3T3 cells expressing 25 different genes.
• Fig. S3. Transforming activity evaluation using the MANO method in vitro.
• Fig. S4. AlamarBlue cell viability assay in Ba/F3 cells expressing EGFR mutants.
• Fig. S5. In vivo evaluation of sensitivity to afatinib using the MANO method.
• Fig. S6. In vitro transforming activity of 101 EGFR mutants evaluated by the MANO method.
• Fig. S7. Immunoblot analysis of Ba/F3 cells expressing EGFR mutants.
• Fig. S8. The droplet digital PCR assay for the detection of EGFR E709A and L858R.
• Fig. S9. Focus-formation assay of 3T3 cells with EGFR L858R compound mutations.
• Fig. S10. Focus-formation assay of 3T3 cells expressing EGFR G719 compound mutations or exon 19 deletion compound mutations.
• Fig. S11. The sensitivity of EGFR mutants to dacomitinib, nazartinib, and neratinib.
• Table S5. Recurrent NSCLC cases after gefitinib treatment without the EGFR T790M mutation.
• Table S7. The frequency and pattern of E709 compound mutations.

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Other Supplementary Material for this manuscript includes the following:

• Table S1. The raw data of Fig. 3A (provided as an Excel file).
• Table S2. One hundred one recurrent EGFR mutations in COSMIC database analyzed with the MANO method (provided as an Excel file).
• Table S3. Barcode sequences used in the MANO method for EGFR compound mutations (provided as an Excel file).
• Table S4. The raw data of Fig. 4C (provided as an Excel file).
• Table S6. Clinical and molecular characteristics of surgically resected lung adenocarcinoma with EGFR compound mutations (provided as an Excel file).
• Table S8. The raw data of Fig. 5C (provided as an Excel file).
• Table S9. The raw data of Fig. 6A (provided as an Excel file).
• Table S10. Candidate drug for each EGFR mutation (provided as an Excel file).

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