|
|
Blood, Vol. 110, Issue 2, 661-669, July 15, 2007
Identification of heat shock protein 32 (Hsp32) as a novel survival factor and therapeutic target in neoplastic mast cells
Blood Kondo et al.
110: 661
Supplemental materials for Kondo et al, Vol. 110, Issue 2, 661-669
Files in this Data Supplement:
- Figure S1. Mast cell culture and development (JPG, 36.7 KB)
-
Mast cells were cultured from their peripheral blood progenitor cells in RPMI 1640 medium plus 10% FCS at 37°C for 21 days, using SCF (100 ng/ml) and IL-6 (100 ng/ml) as growth factors. Effects of various concentrations of SMA-ZnPP on (A) mast cell development (numbers of mast cells) on day 21 and on (B) the percentage of apoptotic cells in the same culture. (C, D) Effects of SMA-ZnPP and PEG-ZnPP on cultured immature MCs on day 21. In this experiment, SCF/IL-6-cultured mast cells on day 21 were exposed to drugs for 48 hours. The number (percentage) of apoptotic cells was determined by light microscopy. Results represent the mean (± SD) of triplicates.
- Figure S2. Effects of PEG-ZnPP and SMA-ZnPP on viability of unstimulated peripheral blood MNC (PB-MNC) from a healthy subject, cultured human umbilical vein endothelial cells (HUVEC), cultured human lung fibroblasts (LUF), and HMC-1 cells (subclones HMC-1.1 and HMC-1.2) as positive control (JPG, 79.8 KB)
-
Cells were cultured in the absence (control) or presence of various concentrations of PEG-ZnPP or SMA-ZnPP as indicated. HUVEC and HMC-1 cells were cultured in IMDM medium, and PB-MNC and LUF were cultured in RPMI 1640 medium. All cells were cultured in 10% FCS. After 48 hours, the numbers (percentages) of apoptotic cells were determined by light microscopy. Visibly, neither PEG-ZnPP nor SMA-ZnPP induced a significant apoptosis in control cells.
- Figure S3. Morphologic properties of cells after exposure to PEG-ZnPP or SMA-ZnPP: photomicrographic examples (JPG, 218 KB)
-
The Hsp32-targeting drugs were applied on peripheral blood MNC (PB-MNC; healthy subject), cultured human umbilical vein endothelial cells (HUVEC), cultured human lung fibroblasts (LUF), and HMC-1 cells (subclones HMC-1.1 and HMC-1.2) at 37°C for 48 hours. HUVEC and HMC-1 cells were cultured in IMDM medium, and PB-MNC and LUF were cultured in RPMI 1640 medium. All cells were cultured in 10% FCS. After 48 hours, cells were spun on cytospin slides and stained by Wright-Giemsa stain. PEG-ZnPP and SMA-ZnPP induced apoptosis in HMC-1 cells, but did not induce apoptosis in PB-MNC, HUVEC, or LUF.a.
|
|
