human monocytic leukemia u937  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Curcuminoids increase global 5hmC in leukemia cells. Graded concentrations of curcumin (Cur) or DMC were used to treat U937 ( a , b ) and HL60 leukemia cells ( c , d ) for 48 h, followed by genomic DNA extraction and quantitation of global 5hmC as described in the methods. The data represent the mean of 3 replicates ± SD. * indicates a significant difference from the control at p < 0.05.

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques: DNA Extraction, Quantitation Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Curcuminoids increase TET enzymatic activity in leukemia cells. Graded concentrations of curcumin (Cur) or DMC were used to treat U937 ( a , b ) and HL60 leukemia cells ( c , d ) for 48 h, followed by nuclear protein extraction and quantitation of TET activity, as described in the methods. The data represent the mean of 3 replicates ± SD. * indicates significant difference from the control at p < 0.05.

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques: Activity Assay, Protein Extraction, Quantitation Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Curcuminoids induce TET isoform transcription in leukemia cells. U937 leukemia cells were treated with graded concentrations of either curcumin for 24 and 48 h ( a ) and ( b ), respectively) or DMC (( c ) and ( d ), respectively), followed by RNA extraction and single-step RT-PCR, as described in the methods. The data represent the mean ± SD for 3 replicates. * indicates a significant difference at p < 0.05, ** indicates a significant difference at p < 0.01, *** indicates a significant difference at p < 0.001, and **** indicates a significant difference at p < 0.0001. Note that TET3 induction is not visible because of the scale.

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques: RNA Extraction, Reverse Transcription Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Volcano plots for curcumin- and DMC-induced changes in 5hmC at the single-CpG level. U937 cells were treated with curcumin (5 µM, ( a )), DMC (1 µM, ( b )), and DMC (2 µM, ( c )) for 48 h, followed by oxidative bisulfite sequencing, as described in the methods. In the volcano plot, each black circle represents a CpG site with a nonsignificant change ( p < 0.05) in 5hmC, while the red circles represent a CpG site with a significant change at p < 0.05. Positive values on the x-axis indicate an increase in 5hmC relative to the control untreated samples, and negative values indicate a decrease in 5hmC relative to the control (active demethylation). The numbers at the top indicate the number of CpG sites showing an increase in DNA 5hmC (positive values on the x-axis) or a decrease in DNA 5hmC (negative values on the x-axis).

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques: Oxidative Bisulfite Sequencing, Control

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Genomic distribution of the increase in 5hmC induced by curcumin and DMC in leukemia cells. U937 leukemia cells treated by curcumin (Cur) (5 μM) or DMC (1 μM) for 48 h, followed by single CpG analysis of 5hmC distribution using RRBS, as described in the methods. The upper panel shows the distribution of the 5hmC increase in promoters, exons, introns, and intergenic regions after treatment with Cur or DMC. The lower panel shows the distribution of 5hmC increase in CpG islands (CpGi), shores, and other regions after treatment with Cur or DMC.

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Curcuminoids Activate TET Enzymes and Increase DNA Hydroxymethylation and Active Demethylation in Leukemia Cells

doi: 10.3390/ijms27010310

Figure Lengend Snippet: Volcano plots for curcumin- and DMC-induced changes in 5hmC at CpG islands and promoter regions. U937 cells were treated with curcumin (5 μM) and DMC (1 μM) for 48 h, followed by oxidative bisulfite sequencing, as described in the methods. For CpG island analysis, ( a , b ) represent curcumin and DMC treatment, respectively. For gene promoter analysis, ( c , d ) represent curcumin and DMC treatment, respectively. In the volcano plot, each black circle represents a CpG site with a nonsignificant change ( p < 0.05) in 5hmC, while the red circles represent a CpG site with a significant change at p < 0.05. Positive values on the x-axis indicate an increase in 5hmC relative to the control untreated samples, and negative values indicate a decrease in 5hmC relative to the control (active demethylation). The numbers at the top indicate the number of CpG sites showing an increase in DNA 5hmC (positive values on the s-axis) or a decrease in DNA 5hmC (negative values on the x-axis).

Article Snippet: Human promyeloid leukemia (HL60) and human monocytic leukemia (U937) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in RPMI-1640 medium (Sigma-Aldrich, Milwaukee, WI, USA) containing 10% fetal bovine serum (FBS) and 2.5 mM L-glutamine.

Techniques: Oxidative Bisulfite Sequencing, Control