Brms1

The HBEC3 cells (Control, p53KD, K-Ras^v12 and HBEC3-p53KD-K-Ras^v12) were a generous gift from John D. Minna at the University of Texas Southwestern Medical Center [17] (link). The HBEC3 cells were immortalized using mouse Cdk4 and hTERT and were maintained in Keratinocyte-SFM (Life Technologies, Grand Island, NY) medium containing 50 µg/mL bovine pituitary extract and 5 ng/mL EGF (Life Technologies). HEK293T cells and NSCLC cell lines (H1975, H1299, A549, H358 and Calu-3) were purchased from ATCC. HEK293T cells were maintained in DMEM with 10% FBS. NSCLC H1975, H1299, A549 and H358 cells were grown in RPMI with 10% FBS. NSCLC Calu-3 cells were maintained in Eagle's Minimum Essential Medium (ATCC) with 10% FBS. Plasmids encoding shRNA BRMS1 or shRNA control were created by inserting the human BRMS1 sequence 5′-gtacatgcttcaagagatc-3′ or a scramble DNA sequence respectively [19] (link) into HpaI-XhoI of the lentiviral pSicoR vector (Addgene, Cambridge, MA). The virus packaging plasmids pMDLg/pRRE, pRSV-Rev and pMD2.G were purchased from Addgene. Primary antibodies used in this study include: p53 and actin (Santa Cruz Biotechnology, Santa Cruz, CA), K-Ras (Abgent, San Diego, CA), BRMS1 (Abcam, Cambridge, MA), and paxillin (Cell Signaling, Danvers, MA). Adenoviral-Cre recombinase (Ad-Cre) was purchased from the University of Iowa (Iowa City, IA).