Journal: Oncotarget

Article Title: Synergistic antitumor activity of histone deacetylase inhibitors and anti-ErbB3 antibody in NSCLC primary cultures via modulation of ErbB receptors expression

doi: 10.18632/oncotarget.7195

Figure Lengend Snippet: Protein lysates from S11 ( A ) and O11 ( B ) cells, untreated or treated for 24 h with vorinostat 1 μM or VPA 0.5 mM and/or A3 25 μg/ml, were immunoblotted with pAKT/AKT, pMAPK/MAPK cleaved PARP and cleaved caspase 8 antibodies. γ-tubulin protein expression was used as protein loading control. ( C ) Cells treated with or without HDACi and/or A3 at the indicated concentration, were collected 24 h after drugs treatment. Then, cells were stained for γH2AX (green) and DAPI for nuclei (blue) and observed by microscope, as indicated in Materials and Methods section.

Article Snippet: Primary antibodies were purchased as follows: EGFR, ErbB2, pErbB3, pAKT, AKT, pMAPK, MAPK, cbl, CHIP, acetyl-H3 and cleaved caspase 8 antibodies from Cell Signaling Technology, Inc. (Boston, MA, USA), ErbB3 and γ-tubulin antibodies from Santa Cruz Biotechnology, Inc. (San Jose, CA, USA), vimentin from DakoCytomation (Glostrup, Denmark), E-cadherin antibody from Abcam (Cambridge, England), cleaved PARP [Poly(ADP-ribose) Polimerase] from BD Pharmingen (Becton Dickinson, San Josè, CA, USA), and γH2AX from Millipore (Billerica, MA, USA).

Techniques: Expressing, Control, Concentration Assay, Staining, Microscopy