1. Iron Chelators Inhibit the Growth and Induce the Apoptosis of Kaposi's Sarcoma Cells and of their Putative Endothelial Precursors 
Thierry Simonart, Michel Heenen 
Journal of Investigative Dermatology 
Volume 115, Issue 5, Pages (November 2000)
DOI: /j x
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Effect of DFO on the expression of proliferation markers by HDMEC, KS spindle cells, and KS-Y1 cells. The cells were exposed to 50 or 200 μM DFO for 32 h. Immunohistochemistry for total PCNA (white columns), stable PCNA (dotted columns), Ki-67 (hatched columns), and cyclin D1 (black columns) was carried out on cytospin preparations, as described in the MaterialS and Method section. Results are expressed as the percentage of positive cells per 500 counted cells. Error bars, SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 significant difference from control.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Immunoblot analysis of p34cdk4 expression in KS-2 and KS-Y1 cells after treatment with DFO. Cell lysates (40 μg of protein for each sample) were separated on a 10–17% sodium dodecyl sulfate–polyacrylamide gel and electrophoretically transferred to nitrocellulose sheets. The proteins were immunoblotted with anti-p34cdk4 (DCS32.2 clone). The immunoblots were scanned with a Microtek Phantom 4800 apparatus. Lane 1, untreated KS-2 cells; lane 2, DFO 50 μM; lane 3, DFO 200 μM; lane 4, untreated KS-Y1 cells; lane 5, DFO 50 μM; lane 6, DFO 200 μM; lane 7, MCF7 cells.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Effect of iron chelation on the terminal deoxynucleotidyl transferase-mediated desoxyuridinetriphosphate nick end labeling-based apoptotic index of HDMEC, KS-2 cells, and KS-Y1 cells. The cells were exposed to the iron chelators for 32 h (white columns) or 72 h (hatched columns). 1, control; 2, DFO 50 μM; 3, deferiprone 150 μM; 4, DFO 200 μM; 5, deferiprone, 450 μM. Results are expressed as the percentage of positive cells per 500 counted cells. No significant difference could be observed between KS-1, KS-2, and KS-3 cells (data not shown). Error bars, SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 significant difference from control.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Morphologic evidence of induction of apoptosis in HDMEC, KS-derived spindle cells, and KS-Y1 cells. Subconfluent cell cultures were treated with DFO 200 μM. After 48 h, the cells were harvested and centrifugated. They were then fixed with 2.5% glutaraldehyde and postfixed for 1 h in 0.1 mM cacodylate-buffered osmium tetroxide. After dehydration in graded series of ethanol, the samples were embedded in Epon 812. One micromolar semithin sections stained with toluidine blue were examined in a Zeiss photomicroscope. (a) HDMEC + DFO 200 μM; (b) HDMEC (control); (c) KS-2 spindle-shaped cells + DFO 200 μM; (d) KS-2 spindle-shaped cells (control); (e) KS-Y1 cells + DFO 200 μM; (f) KS-Y1 cells (control)
Journal of Investigative Dermatology  , DOI: ( /j x)
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6. Figure 5
Viability assays of KS-derived cells, KS Y-1 cells and HDMEC during serum deprivation, as determined by the trypan blue exclusion test. Cultures were harvested at day 1 (white columns), day 3 (hatched columns), and day 7 (black columns) after serum deprivation. The cells were suspended in PBS, mixed with Trypan blue and incubated for 3 min. Direct counting of cells excluding and taking up the dye was performed with a hemocytometer. Cell counts are indicated as the percentage of viable cell numbers at day 0. Effect of (a) serum deprivation alone; (b) serum deprivation + FeCl3 20 μM; (c) serum deprivation + DFO 20 μM. No significant difference could be observed between KS-1, KS-2, and KS-3 cells (data not shown). Error bars, SEM (n = 3).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Immunoblot analysis of Bcl-2 expression in KS-2 and KS-Y1 cells after treatment with DFO. Cell lysates (40 μg of protein for each sample) were separated on a 10–17% sodium dodecyl sulfate–polyacrylamide gel and electrophoretically transferred to nitrocellulose sheets. The proteins were immunoblotted with anti-Bcl-2 (100 clone). The immunoblots were scanned with a Microtek Phantom 4800 apparatus. Lane 1, untreated KS-2 cells; lane 2, DFO 100 μM; lane 3, DFO 250 μM; lane 4, untreated KS-Y1 cells; lane 5, DFO 100 μM; lane 6, DFO 250 μM.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions