Journal: British Journal of Cancer

Article Title: Co-targeting of FAK and MDM2 triggers additive anti-proliferative effects in mesothelioma via a coordinated reactivation of p53

doi: 10.1038/bjc.2016.331

Figure Lengend Snippet: NF2 regulates the interaction of FAK–p53 and MDM2–p53. ( A ) The NF2–FAK complex was evaluated in MESO257 by NF2 and FAK immunoprecipitation followed by FAK and NF2 immunoblotting. ( B ) Nuclear localisation of NF2, FAK, p53, and MDM2 was evaluated in MESO257 by immunoblotting. Poly(ADP-ribose) polymerase (PARP) is a nuclear localisation control, and GAPDH is a cytoplasmic control. ( C ) In MESO257 with stable NF2 shNRA expression, p53, NF2, FAK, MDM2, and p21 expression were evaluated by immunoblotting after FAK shRNA knockdown for 72 h. β -Actin staining is a loading control. p53 expression quantifications are standardised to the empty vector control. ( D ) Expression of p53 and p21 was evaluated in MESO257 cell membrane, cytoplasm, and nucleus with stable NF2 shNRA expression by immunoblotting. ( E ) Cell viability was evaluated by a cell titre Glo ATP-based luminescence assay in MESO257 with stable expressed NF2 shRNA , at 72 h post-infection with FAK shRNA . Data were normalised to empty lentivirus infections, and represent the mean values (±s.d.) from quadruplicate cultures. Statistically significant differences between untreated control and treatments or between vector control and FAK shRNA or NF2 shRNA and NF2+FAK shRNA are presented as * P <0.05, ** P <0.01.

Article Snippet: Human NF2 full-length cDNA expression plasmid (Catalogue: TC124024) was obtained from Origene (Rockville, MD, USA).

Techniques: Immunoprecipitation, Western Blot, Control, Expressing, shRNA, Knockdown, Staining, Plasmid Preparation, Membrane, Luminescence Assay, Infection