fig3
Figure 3. HF induces resistance to apoptosis in AML cells. (A) Representative cytograms of AML blood cells treated for 72 h with HF (1.4, 2, and 3 µg/mL); detection of dead cells after annexin-V-FITC/PI staining and flow cytometry. The percentage of annexin-V-positive cells is shown. (B) Light microscopy of non-treated and AML cells treated with HF: original magnification, 600×; May-Grünwald stain; Scale bar, 10 µm. (C) Representative cytograms of PBMCs and isolated monocytes from two healthy donors, treated with 2 µg/mL HF for 72 h, and cell death was assessed as described in (A). (D) Representative cytograms of AML cells isolated from blood and bone marrow of one AML patient, treated with 2 µg/mL HF for 72 h, and cell death was assessed as described in (A). (E) Cell death levels (2 µg/mL, 72 h) were determined in untreated and HF-treated malignant cells from 35 AML patients. Mean concentrations ± SD are indicated; P value was calculated using a Mann-Whitney U-test; ****P < 0.0001. (F) The percentage of HF-mediated AML cell death was determined for all FAB subtypes by subtracting the percentage of annexin-V-positive cells in the absence of HF from the percentage of annexin-positive cells in the presence of HF, and then dividing by the percentage of annexin-positive cells in the presence of HF × 100. Data are the mean ± SEM (M0, n = 3; M1, n = 8; M2, n = 14; M3, n = 1; M4, n = 5; and M5, n = 4). P values were calculated using one-way ANOVA (without M3 value); ns: not significant. (G&H) AML cells were treated with HF
