Journal: Cancer Biology & Therapy

Article Title: EGFR signaling defines Mcl - 1 survival dependency in neuroblastoma

doi: 10.1080/15384047.2014.1002333

Figure Lengend Snippet: Mcl-1 dependent NB cells NLF (A) and SK-N-BE(2) (B) were exposed to EGFR inhibitors, erlotinib or cetuximab, and ABT-737 simultaneously and evaluated after 48 hours for changes in survival by WST-1. (C) NLF cells were exposed to given concentrations of U0126, erlotinib, or LY294002 for 24 hours, harvested for protein, and evaluated for changes in EGFR signaling and Bcl-2 family protein expression. (D) Protein from an Mcl-1 co-IP was treated with Lambda Protein Phosphatase (LPP) and then evaluated for changes in Bim:Mcl-1 interactions by immunoblot.

Article Snippet: 4 The Mcl - 1 protein;Mcl - 1 antibody matrix was then pelleted with centrifugation, and resuspended in a lambda phosphatase (LPP, Promega, Sweden) containing solution (4 uL LPP:4 uL 10X Buffer: 32 uL co-IP solution + 1 M DTT (Millipore)) and incubated at 37°C for 40 min. An equal portion of Mcl - 1 protein;Mcl - 1 antibody matrix was resuspended in IP buffer alone (without LPP) and incubated at 37°C for 40 min to control for the effects of heating on protein interactions.

Techniques: Expressing, Co-Immunoprecipitation Assay, Western Blot