Fig. 3.
Fig. 3. Influence of the cellular context and exposure schedule on the potentiation of the ATRA-dependent induction of NBT-reducing activity by ST1346. / (A) NB4 cells were treated for 3 days with vehicle (○) or ST1346 (5 μM) (●) in the presence of the indicated concentrations of ATRA. (B) NB4 cells were treated with vehicle, ST1346 (5 μM), ATRA (0.1 μM), or the combination of ATRA + ST1346 (ST1346/ATRA) for the indicated amount of time in days (d). Pretreatment means that ST1346 or ATRA (+) is added to the culture medium for the indicated amount of time. This is followed by a cell-washing step in complete culture medium and subsequent addition of fresh medium containing the complementary compound not present in the pretreatment phase. In the case of ATRA + ST1346, we added a control group in which a washing step was introduced after 3 days of pretreatment (rightmost data bar). (D) PR9 cells were treated with ATRA (1 μM), ST1346 (10 μM), or the combination of the 2 compounds for 3 days in the absence or presence of the PML-RARα transgene inducer zinc sulfate (Zn++; 50 μM). (C,E-H) NB4, NB4.306, HL-60, U937, Kazumi, or KG1 cells were treated with vehicle (control), ST1346 (at the indicated concentrations), ATRA (0.1 or 1 μM), or the combination of ATRA + ST1346 for 3 days (panels C, E, F, and H) or the indicated number of days (panel G). For all the experiments, cells were seeded at a concentration of 200 000/mL and aliquots used for the determination of NBT-reducing activity (panels A, B, C, and E) or the number of NBT+ cells (panels D, F, G, and H). In all cases the cellular viability was above 90%, as assessed by the erythrosin exclusion test. The results are the means ± SD of 3 separate cultures. *Significantly higher than the corresponding group treated with vehicle (P < .01 according to the Studentt test). °Significantly higher than the sum of the effects observed in the corresponding ST1346- and ATRA-treated groups, following 2-way analysis of variance and measurement of the F of interaction (P < .01 according to Tukey test).

Influence of the cellular context and exposure schedule on the potentiation of the ATRA-dependent induction of NBT-reducing activity by ST1346.

(A) NB4 cells were treated for 3 days with vehicle (○) or ST1346 (5 μM) (●) in the presence of the indicated concentrations of ATRA. (B) NB4 cells were treated with vehicle, ST1346 (5 μM), ATRA (0.1 μM), or the combination of ATRA + ST1346 (ST1346/ATRA) for the indicated amount of time in days (d). Pretreatment means that ST1346 or ATRA (+) is added to the culture medium for the indicated amount of time. This is followed by a cell-washing step in complete culture medium and subsequent addition of fresh medium containing the complementary compound not present in the pretreatment phase. In the case of ATRA + ST1346, we added a control group in which a washing step was introduced after 3 days of pretreatment (rightmost data bar). (D) PR9 cells were treated with ATRA (1 μM), ST1346 (10 μM), or the combination of the 2 compounds for 3 days in the absence or presence of the PML-RARα transgene inducer zinc sulfate (Zn++; 50 μM). (C,E-H) NB4, NB4.306, HL-60, U937, Kazumi, or KG1 cells were treated with vehicle (control), ST1346 (at the indicated concentrations), ATRA (0.1 or 1 μM), or the combination of ATRA + ST1346 for 3 days (panels C, E, F, and H) or the indicated number of days (panel G). For all the experiments, cells were seeded at a concentration of 200 000/mL and aliquots used for the determination of NBT-reducing activity (panels A, B, C, and E) or the number of NBT+ cells (panels D, F, G, and H). In all cases the cellular viability was above 90%, as assessed by the erythrosin exclusion test. The results are the means ± SD of 3 separate cultures. *Significantly higher than the corresponding group treated with vehicle (P < .01 according to the Studentt test). °Significantly higher than the sum of the effects observed in the corresponding ST1346- and ATRA-treated groups, following 2-way analysis of variance and measurement of the F of interaction (P < .01 according to Tukey test).

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