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. Author manuscript; available in PMC: 2021 May 1.
Published in final edited form as: Mol Cancer Res. 2020 Aug 14;18(11):1675–1684. doi: 10.1158/1541-7786.MCR-20-0262

Figure 1. CRISPR Synergistic Activation Mediator (SAM) system can be used to model RICTOR amplification in NSCLC.

Figure 1.

(A-C) Patient data from the TCGA Pan-Lung Cancer Atlas were analyzed using the online platform at cbioportal.org. (A) Alteration frequencies of RICTOR in non-small cell lung cancer, squamous cell carcinoma, and adenocarcinoma. (B) Alterations in mTOR complex components in patient-matched samples. (C) Correlation of RICTOR mRNA levels with copy number values in Lung Adenocarcinoma. Red, amplified samples based on GISTIC scoring. (D-H) CRISPR SAM (15) was used to target the promoter region of Rictor 100 bp (R5) and 121 bp (R6) upstream of the transcription start site (TSS). (D) in a panel of immortalized lung epithelium and NSCLC cell lines. (E) RICTOR copy number values of utilized cell lines according to CCLE database. (F) Western blot analysis of Rictor protein levels in parental cell lines. mRNA relative to Actin (G) and protein levels of Rictor (H) were measured by qRT-PCR or Western blotting, respectively. Quantification of Rictor expression normalized to tubulin is indicated in numbers below the blots. EV: empty vector