NuRD and HP1γ inactivation phenocopy H9SB at the functional and transcriptional level. (A) Growth kinetics of MOLM13 cells treated with panobinostat (Pano; 4 nM), chaetocin (Ch) (40 nM) alone, or in combination. The data are shown as the averages of biological replicates (n = 3) ± SD. Two-way ANOVA test, ∗P < .01 comparing NT vs treatments at 72 hours. (B) The bar graph shows the differentiation measured with CD11b-CD15 surface expression in MOLM13 cells after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination, over the time course. Data shown are the averages of 3 biological replicates ± SD. Two-way ANOVA test, ∗P < .001. (C) The histogram shows flow cytometric analyses of annexin V–positive MOLM13 cells 72 hours after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination. Representative plots of 3 independent biological replicates are shown. (D) qRT-PCR expression levels of selected target genes in MOLM13 cells treated with drugs alone or in combination for 48 hours. The data shown are representative of 3 independent biological replicates. (E) Growth kinetics of OCIAML3 cells treated with Pano (4 nM), Ch (40 nM) alone, or their combination. Fifty thousand cells were seeded followed by daily counting. The data are shown as the averages of 3 biological replicates ± SD. Statistical significance calculated using the 2-way ANOVA test, ∗P < .01. (F) The bar graph shows differentiation measured by CD11b-CD15 surface expression in OCIAML3 cells after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination, for the time course. Data shown are the averages of 3 biological replicates ± SD. Statistical significance calculated using the 2-way ANOVA test, ∗∗P < .001. (G) The histogram shows flow cytometric analyses of annexin V–positive OCIAML3 cells 72 hours after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination. Representative plots of 3 independent biological replicates are shown. (H) qRT-PCR expression levels of selected target genes in OCIAML3 cells treated with drugs alone or in combination for 48 hours. The data shown here are representative of 3 independent biological replicates. (I) Percent viability determined via annexin V/7AAD staining in primary AML cells after treatment with a combination of Pano (4 nM) + Ch (40 nM) or dimethyl sulfoxide (DMSO). (J) qRT-PCR expression levels of selected target genes in primary AML cells treated with drugs in combination or DMSO for 48 hours. The data shown here are the averages of triplicates of quantitative PCR values.

NuRD and HP1γ inactivation phenocopy H9SB at the functional and transcriptional level. (A) Growth kinetics of MOLM13 cells treated with panobinostat (Pano; 4 nM), chaetocin (Ch) (40 nM) alone, or in combination. The data are shown as the averages of biological replicates (n = 3) ± SD. Two-way ANOVA test, ∗P < .01 comparing NT vs treatments at 72 hours. (B) The bar graph shows the differentiation measured with CD11b-CD15 surface expression in MOLM13 cells after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination, over the time course. Data shown are the averages of 3 biological replicates ± SD. Two-way ANOVA test, ∗P < .001. (C) The histogram shows flow cytometric analyses of annexin V–positive MOLM13 cells 72 hours after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination. Representative plots of 3 independent biological replicates are shown. (D) qRT-PCR expression levels of selected target genes in MOLM13 cells treated with drugs alone or in combination for 48 hours. The data shown are representative of 3 independent biological replicates. (E) Growth kinetics of OCIAML3 cells treated with Pano (4 nM), Ch (40 nM) alone, or their combination. Fifty thousand cells were seeded followed by daily counting. The data are shown as the averages of 3 biological replicates ± SD. Statistical significance calculated using the 2-way ANOVA test, ∗P < .01. (F) The bar graph shows differentiation measured by CD11b-CD15 surface expression in OCIAML3 cells after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination, for the time course. Data shown are the averages of 3 biological replicates ± SD. Statistical significance calculated using the 2-way ANOVA test, ∗∗P < .001. (G) The histogram shows flow cytometric analyses of annexin V–positive OCIAML3 cells 72 hours after treatment with Pano (4 nM), Ch (40 nM) alone, or their combination. Representative plots of 3 independent biological replicates are shown. (H) qRT-PCR expression levels of selected target genes in OCIAML3 cells treated with drugs alone or in combination for 48 hours. The data shown here are representative of 3 independent biological replicates. (I) Percent viability determined via annexin V/7AAD staining in primary AML cells after treatment with a combination of Pano (4 nM) + Ch (40 nM) or dimethyl sulfoxide (DMSO). (J) qRT-PCR expression levels of selected target genes in primary AML cells treated with drugs in combination or DMSO for 48 hours. The data shown here are the averages of triplicates of quantitative PCR values.

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