Journal: Oncotarget

Article Title: HoxA9 regulated Bcl-2 expression mediates survival of myeloid progenitors and the severity of HoxA9-dependent leukemia

doi:

Figure Lengend Snippet: (A) The viability of wild type and Bax;Bak −/− HoxA9 FDM cells was determined by flow cytometry analysis of AnnexinV-FITC staining and PI exclusion, at 0, 4, 8 and 12 days following removal of 4-OHT. Results are means ±SEM of 2 pools and 2 clones repeated in 3 independent experiments. (B) Bax;Bak −/− HoxA9 immortalized cells were cultured in the presence (Day 0) or absence of 4-OHT over 6 days. At the indicated times, lysates were harvested and Western blots probed with antibodies against HoxA9, Bcl-2, Mcl-1 and Bcl-x L . Actin was used as a loading control.

Article Snippet: The antibodies used were; Anti-mouse Bcl-2 mAb (BD Biosciences), anti-Actin mAb and pAb anti-Bax and anti-Bak (Sigma), anti-Bad pAb (Cell Signaling), anti-Bim pAb (Stressgen), anti-Puma pAb (ProSci), anti-HoxA9 pAb and anti-MLL clone N4.4 mAb (Millipore), anti-Mcl-1 pAb (Rockland) and anti-Bcl-x L pAb (R&D Systems). pAb anti-Bid, anti-Bmf and anti-A1 pAb were a kind gift from Andreas Strasser, Lorraine O'Reilly and Marco Herold respectively.

Techniques: Flow Cytometry, Staining, Clone Assay, Cell Culture, Western Blot