Figure 5
Figure 5. SMA-ZnPP induces apoptosis in CML cells. (A) K562 cells were incubated with control medium (left panel) or SMA-ZnPP (10 μM) (right panel) for 48 hours. Thereafter, cells were analyzed by electron microscopy as described in “Methods.” SMA-ZnPP was found to induce typical morphological signs of apoptosis (chromatin fragmentation/condensation). (B) K562 cells were incubated with control medium (left panel) or SMA-ZnPP (10 μM) (right panel) for 48 hours. Thereafter, cells were subjected to a Tunel assay. Apoptotic cells display bright green fluorescence. (C) K562 cells were incubated in control medium (Co) or in PEG-ZnPP or SMA-ZnPP (each 10 or 20 μM) at 37°C. After 48 hours, cells were subjected to annexin-V staining. Results show the percentage of annexin-V–positive cells and represent the mean ± SD of 3 independent experiments. *P < .05 compared with control. (D) K562-KRAB-HO-1 cells were cultured in control medium (Co) or in the presence of doxycycline (1 μg/ml) for 24 hours. Thereafter, expression of Hsp32/HO-1 was determined by Western blotting. β-actin is shown as loading control. (E) K562-KRAB-HO-1 cells were cultured in control medium (Co) or in the presence of doxycycline (1 μg/mL) for 24 hours. Thereafter, cells were kept in the absence or presence of SMA-ZnPP (10 μM) as indicated for 48 hours. Cell viability was determined by trypan blue exclusion test. Results show the percentage of trypan blue-positive cells and represent the mean (± SD) of 3 independent experiments. (F) Leukemic cells from untreated patients with CP CML were cultured in control medium or in the presence of SMA-ZnPP (10 μM or 30 μM) for 4 days. Thereafter, the percentage of apoptotic cells was determined by annexin-V staining and FACS analysis. Results represent the mean (± SD) of 3 patients. *P < .05. (G) Primary CML cells were cultured in the absence or presence of SMA-ZnPP (10 μM) for 5 days. Thereafter, the percentage of apoptotic cells was quantified on Wright-Giemsa-stained cytospin preparations. Results represent the mean (± SD) of 3 patients. *P < .05.

SMA-ZnPP induces apoptosis in CML cells. (A) K562 cells were incubated with control medium (left panel) or SMA-ZnPP (10 μM) (right panel) for 48 hours. Thereafter, cells were analyzed by electron microscopy as described in “Methods.” SMA-ZnPP was found to induce typical morphological signs of apoptosis (chromatin fragmentation/condensation). (B) K562 cells were incubated with control medium (left panel) or SMA-ZnPP (10 μM) (right panel) for 48 hours. Thereafter, cells were subjected to a Tunel assay. Apoptotic cells display bright green fluorescence. (C) K562 cells were incubated in control medium (Co) or in PEG-ZnPP or SMA-ZnPP (each 10 or 20 μM) at 37°C. After 48 hours, cells were subjected to annexin-V staining. Results show the percentage of annexin-V–positive cells and represent the mean ± SD of 3 independent experiments. *P < .05 compared with control. (D) K562-KRAB-HO-1 cells were cultured in control medium (Co) or in the presence of doxycycline (1 μg/ml) for 24 hours. Thereafter, expression of Hsp32/HO-1 was determined by Western blotting. β-actin is shown as loading control. (E) K562-KRAB-HO-1 cells were cultured in control medium (Co) or in the presence of doxycycline (1 μg/mL) for 24 hours. Thereafter, cells were kept in the absence or presence of SMA-ZnPP (10 μM) as indicated for 48 hours. Cell viability was determined by trypan blue exclusion test. Results show the percentage of trypan blue-positive cells and represent the mean (± SD) of 3 independent experiments. (F) Leukemic cells from untreated patients with CP CML were cultured in control medium or in the presence of SMA-ZnPP (10 μM or 30 μM) for 4 days. Thereafter, the percentage of apoptotic cells was determined by annexin-V staining and FACS analysis. Results represent the mean (± SD) of 3 patients. *P < .05. (G) Primary CML cells were cultured in the absence or presence of SMA-ZnPP (10 μM) for 5 days. Thereafter, the percentage of apoptotic cells was quantified on Wright-Giemsa-stained cytospin preparations. Results represent the mean (± SD) of 3 patients. *P < .05.

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