|
|
Blood, Vol. 108, Issue 5, 1461-1468, September 1, 2006
Cell death provoked by loss of interleukin-3 signaling is independent of Bad, Bim, and PI3 kinase, but depends in part on Puma
Blood Ekert et al.
108: 1461
Supplemental materials for: Ekert et al, Vol 108, Issue 5, 1461-1468
Files in this Data Supplement:
- Document 1. Methods (PDF, 23 KB)
- Figure S1. Survival following IL-3 withdrawal in Bid–/–, Hrk–/–, and Puma–/–;Noxa–/– cells (JPG, 51.7 KB)
-
(A) Multiple independent cell lines lacking Bid (n = 3), Hrk (n = 2), both Puma and Noxa (n = 3), only Puma (n = 3), both Bax and Bak (n = 2), or wild-type cells (n = 2) were starved of IL-3 over a 5-day time course and cell viability determined by staining with FITC-coupled annexin V and PI followed by flow cytometric analysis. (B) Lysates of wild-type and Bax–/–;Bak–/– FDM cells cultured in the presence or absence of IL-3 for 0, 6, 24, or 48 hours were immunoblotted with antibodies against Bid or Mcl-1. Probing with an antibody to  -actin was used as a loading control. Lysates from FDC-P1 cells cultured in the presence or absence of IL-3 for 8 hours and Bid–/– cells cultured in the presence of IL-3 are shown as controls.
- Figure S2. Treatment with wortmannin enhances IL-3 withdrawal–induced apoptosis (JPG, 66.3 KB)
-
FDM cells of the indicated genotypes were cultured in the presence or absence of IL-3 for 24 hours with or without wortmannin at 0 µM, 0.5 µM, or 5.0 µM. Cell viability was determined by staining with FITC-coupled annexin V and PI followed by flow cytometric analysis. Results represent the mean ± SE of 3 independent experiments.
|
|
