|
|
Blood, Vol. 111, Issue 6, 3211-3219, March 15, 2008
Bcl-2 antagonist apogossypol (NSC736630) displays single-agent activity in Bcl-2–transgenic mice and has superior efficacy with less toxicity compared with gossypol (NSC19048)
Blood Kitada et al.
111: 3211
Supplemental materials for: Kitada et al
Files in this Data Supplement:
- Figure S1. Toxicity analysis of Gossypol and ApoGossypol (JPG, 54.3 KB)
-
Balb/c mice were orally administered vehicle control ( C ), Gossypol (G), or ApoGossypol (A) at 120 µmol/kg 5-times weekly for 3 weeks. Serum levels of BUN (A) and Creatinine (B) were measured by the use of an automated blood chemistry analyzer COBAS MIRA Classic. Hematological profiles were analyzed using peripheral blood for white blood cells (C), red blood cells (D), lymphocytes (E), and platelets (F) by the use of an automated hematology analyzer VetScan HM2. All data represent mean ± SD of n = 6 per group of mice.
- Figure S2. Time-course of killing of NHL cell line by ApoGossypol (JPG, 21.5 KB)
-
DoHH2 cells were cultured for various times with 10 µM ApoGossypol. %Viable cells was determined by FITC-annexin/PI 2-color staining using a 3-color FACSort instrument and FACS data were analyzed by the use of Flow-Jo Program.
- Figure S3. Effects of ApoGossypol on splenic B-cells (JPG, 37.9 KB)
-
Splenocytes from a Bcl-2 transgenic mouse were isolated, cultured for 18 hrs with various concentrations of ApoGossypol, and 4-color FACS analysis was performed using PE-conjugated anti-B220 antibody in combination with FITC-Annexin V, PI and APC-conjugated CD3 antibody to determine the percentage of apoptotic B-cells (Annexin+, PI−, B220+) among all B220+ cells via 6-color FACSCanto instrument, and FACS data were analyzed by the use of Flow-Jo Program.
- Figure S4. Effect of culture cell density on sensitivity of Bcl-2-transgenic splenocytes to ApoGossypol and Gossypol (JPG, 51.4 KB)
-
Splenocytes from two Bcl-2 transgenic mice (designated mouse A and mouse B) were isolated and cultured at 107 cells/mL for 18 hrs with various concentrations log-scale of either ApoGossypol (A) or Gossypol (G). %Viability was determined by FITC-Annexin V/PI staining, scoring as viable the Annexin V-/PI- cells by FACS analysis.
- Figure S5. Effects of ApoGossypol and Gossypol on Bax/Bak deficient murine fibroblasts (JPG, 35.6 KB)
-
Wild type (WT) and Bax/Bak double knock-out (DKO) mouse embryonic fibroblasts (MEF) were cultured in 24-well plates with various concentrations (0-10 µM) of ApoGossypol or Gossypol. After ∼18-hours, cells were recovered, including floating cells and attached cells (after brief incubation with 0.05% trypsin-EDTA), and stained with FITC-Annexin V plus propidium iodide (PI), then analyzed by FACS. Data represent % viable cells (Annexin V-negative/PI-negative) measured from duplicate determinations (mean ± std error).
|
|
