Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: H2O2-induced different cell viability in M17, PC12 and MN9D cells as measured by MTT assay (A & B) and trypan blue dye exclusion assay (C). Each bar from Figs. represents data obtained from 5 separate experiments (N = 5 of independent cell culture preparations, which is the same in the following legends). *p < 0.05, **p < 0.01, ***p < 0.001, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: MTT Assay, Exclusion Assay, Cell Culture, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on DDRs in M17 and MN9D cells as demonstrated by measurements of γH2AX (A) and p53 (B). Cells were exposed to 100–400 μM H2O2 for 3 h. Top panels: autoradiograph obtained by western blotting. Button panel: the quantitative analysis of band densities in western blotting. Each bar from Figs. represents data obtained from 5–7 separate experiments (N = 5–7). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Autoradiography, Western Blot, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: H2O2 treatment induces single-strand DNA breaks as determined by the Comet assay in M17 and MN9D cells. Cells were exposed to different concentrations of H2O2 for 3 h. The cells were processed for comet assays run under alkaline conditions to identify DNA SSBs. Tail moment (B), tail length (C) and % DNA in the tail (D) were analyzed to evaluate DNA damage. Each bar from Figs. represents data obtained from 5 separate experiments (N = 5). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Single Cell Gel Electrophoresis, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on ROS production in M17 and MN9D cells. Cells were exposed to different concentrations of for 3 h. ROS was measured by the DCFH2-DA assay. Each bar from Figs. represents data obtained from 6 separate experiments (N = 6). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on Ctr1 (A) and OGG1 (B) protein levels in M17 and MN9D cells. Each bar represents data obtained from 5 separate experiments (N = 5) in (A), and 4 separate experiments (N = 4) in (B). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on Cav1.2 (A) and Cav1.3 (B) protein levels in M17 and MN9D cells as determined by western blotting. Cells were exposed to different concentrations of H2O2 for 3 h. Each bar in Figs. represents data obtained from 6 to 7 separate experiments (N = 6–7). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Western Blot, Control

Journal: Journal of immunotoxicology

Article Title: The early growth response factor-1 contributes to interleukin-13 production by mast cells in response to stem cell factor stimulation.

doi: 10.1080/15476910802129612

Figure Lengend Snippet: FIG. 4. Egr-1 deficiency has no effects on c-Kit and IgE receptor expression or mast-cell viability. (A) Bone marrow cells from wild-type mice or Egr-1– deficient mice were cultured in conditioned media in vitro for 4 wk and exam- ined by flow cytometry for c-Kit and IgE receptor expression. For c-Kit anal- ysis, BMMCs were stained with FITC conjugated rat anti-mouse c-Kit mAb or FITC conjugated rat IgG2a isotypic control. For analysis of IgE receptor expression, BMMC were sensitized with IgE overnight and then stained with FITC-conjugated anti-IgE antibody (mouse IgG1). No difference in c-Kit or IgE receptor expression was observed between Egr-1+/+ and Egr-1−/−BMMCs. (B) BMMC from Egr-1-deficient mice or wild-type mice cultured in complete media containing WEHI-3B supernatants (a source of IL-3) were healthy and showed ≥96% viability. To induce mast cell death, WEHI-3B supernatant was removed from culture media for various days. Mast cell viability was examined by trypan blue exclusion assay. Data were expressed as mean ± SD (n = 3 independent experiments).

Article Snippet: Fluorescein isothiocyanate (FITC) anti-mouse CD117 monoclonal antibody (mAb) (CL8936F), FITC rat IgG2a (CLCR2A01) were purchased from Cedarlane Laboratories Limited (Ontario, Canada).

Techniques: Expressing, Cell Culture, In Vitro, Cytometry, Staining, Control, Trypan Blue Exclusion Assay

Journal: Molecular medicine reports

Article Title: SH003 selectively induces p73‑dependent apoptosis in triple‑negative breast cancer cells.

doi: 10.3892/mmr.2016.5722

Figure Lengend Snippet: Figure 1. SH003 selectively inhibits the cell viability of TNBC cells. (A) TNBC cell lines (Hs578T, MDA‑MB231 and ZR‑75‑1) and non‑TNBC cell lines (MCF‑7 and T47D) were treated with the indicated concentrations of SH003, and their cell numbers were assessed by a trypan blue exclusion assay. Data are presented as the mean ± standard deviation (n=6). *P<0.05 and **P<0.01, compared with the untreated cells. (B) MDA‑MB‑231 and MCF‑7 cells were pre‑treated with the indicated concentrations of SH003 for 24 h. After incubation for 15 days, colonies with >50 cells were counted. Data are presented as the mean ± standard deviation (n=3). *P<0.005 compared with the non‑treated cells. TNBC, triple‑negative breast cancer.

Article Snippet: Established Hs578T, MDA-MB-231, ZR-75-1, MCF7 and T47D human breast cancer cell lines were purchased from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Trypan Blue Exclusion Assay, Standard Deviation, Incubation

Journal: Journal of Biological Chemistry

Article Title: Improved Pharmacokinetics of Recombinant Bispecific Antibody Molecules by Fusion to Human Serum Albumin

doi: 10.1074/jbc.m700820200

Figure Lengend Snippet: FIGURE 5. Binding of recombinant antibodies and antibody-HSA fusion proteins to CEA in ELISA. Titration of scDb (A), scFvCEA (B), and taFv (C) and the respective HSA fusion proteins for binding to CEA. Detection was per- formed with an HRP-conjugated anti-His tag antibody. Duplicate samples were analyzed.

Article Snippet: Anti-human IL-2 antibodies as well as the standard of recombinant human IL-2 was provided by the DuoSet IL-2 ELISA Development System kit (R&D Systems, Nordenstadt, Germany) and the assay was performed following the manufacturer’s protocol.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Titration

Journal: Journal of Biological Chemistry

Article Title: Improved Pharmacokinetics of Recombinant Bispecific Antibody Molecules by Fusion to Human Serum Albumin

doi: 10.1074/jbc.m700820200

Figure Lengend Snippet: FIGURE 6. Characterization of B7-DbCEA. A, Coomassie-stained SDS-PAGE (10%) of purified B7-DbCEA pro- tein under reducing and non-reducing conditions. B, size exclusion chromatography of B7-DbCEA by HPLC. C, simultaneous binding of B7-DbCEA to CEA and recombinant CTLA-4. ELISA plate was coated with CEA (3 g/ml) and incubated with 1 M B7-DbCEA or DbCEA. Detection was performed by CTLA-4-Fc, followed by anti-mouse Fc-HRP. D, costimulatory property of B7-Db. PBMCs were incubated for 22 h with 15 nM scDb- CEACD3 in the presence or absence of B7-DbCEA in solution or with the constructs presented on LS174T cells. IL-2 release was determined by ELISA from duplicate samples.

Article Snippet: Anti-human IL-2 antibodies as well as the standard of recombinant human IL-2 was provided by the DuoSet IL-2 ELISA Development System kit (R&D Systems, Nordenstadt, Germany) and the assay was performed following the manufacturer’s protocol.

Techniques: Staining, SDS Page, Purification, Size-exclusion Chromatography, Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Incubation, Construct

Journal: Journal of Biological Chemistry

Article Title: Improved Pharmacokinetics of Recombinant Bispecific Antibody Molecules by Fusion to Human Serum Albumin

doi: 10.1074/jbc.m700820200

Figure Lengend Snippet: FIGURE 7. Target cell-mediated activation of PBMCs by bispecific recombinant antibody-HSA fusion proteins. LS174T (A) and HT1080#13.8 (B) cells were preincubated for 1 h with 40 nM bispecific recombi- nant antibody construct in the presence (B7-DbCEA) or absence (scFvCEA) of 40 nM costimulus. PBMCs were added and IL-2 concentration in the cell culture supernatant measured after 22–24 h by ELISA from duplicate samples. Antibody constructs were further analyzed on LS174T cells applying 5 (C) or 0.2 (D) nM antibody molecules.

Article Snippet: Anti-human IL-2 antibodies as well as the standard of recombinant human IL-2 was provided by the DuoSet IL-2 ELISA Development System kit (R&D Systems, Nordenstadt, Germany) and the assay was performed following the manufacturer’s protocol.

Techniques: Activation Assay, Recombinant, Construct, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A KO in APL cell line NB4 prohibited proliferation, colony formation, and induced differentiation. (A) Proliferation assay of NB4 cells transduced with 2 control sgRNAs (Ctrl) and 2 KDM5A sgRNAs (KDM5A KO). Cell counts were measured at the indicated days after seeding. Data are shown as means ± standard error of the mean of triplicate analysis. The counts of KDM5A-KO cell lines at day 4 were compared with those in control cell lines. Statistical significance was determined by two-way analysis of variance. ****P < .0001. The P values of both KDM5A-KO groups were < .0001. (B-C) Bar graph and photos showing colony formation of the indicated NB4 cell lines using a 14-day methylcellulose assay. Colony counts were compared between 2 control cell lines and 2 KDM5A-KO cell lines, and statistical significance was determined by two-way analysis of variance. ***P < .001. (D) RT-qPCR analysis of differentiation markers CD11b, CD11c, and CD14 in the indicated cell lines. (E-F) Differentiation marker CD11b level in NB4 cells transduced with control sgRNAs and KDM5A sgRNAs measured by using flow cytometry. The statistical significance of RT-qPCR and percentage of CD11b+ cells was determined by two-way analysis of variance. ***P < .001. FACS, fluorescence-activated cell sorting.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Proliferation Assay, Transduction, Methylcellulose Assay, Quantitative RT-PCR, Marker, Flow Cytometry, Fluorescence, FACS

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A KO led to the elevation of H3K4me2 of KDM5A target genes. (A) Pie chart showing the genomic distribution of KDM5A ChIP-seq peaks, annotated by using HOMER. The numbers represent the peak number in each category. (B) Heatmap analysis of the ChIP-seq profiles of KDM5A in NB4 and H3K4me2 and H3K4me3 in the control (Ctrl) and KDM5A-KO NB4 cells at KDM5A bound genes. Gene bodies ± 10 kb are shown, sorted by the intensity of KDM5A binding at promoter regions. (C) ChIP-seq signal density plots showing the global levels of H3K4me3 and H3K4me2 at KDM5A bound genes ± 10 kb. TES, transcription end site; TSS, transcription start site.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: ChIP-sequencing, Binding Assay

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A KO led to the increase of H3K4me2 at promoters and 5' intragenic regions correlated with upregulation of gene expression. (A) Heatmap analysis of the ChIP-seq profiles of KDM5A in NB4 and H3K4me2 and H3K4me3 in the control (Ctrl) and KDM5A-KO NB4 cells. Gene bodies with upregulated H3K4me2 (in KDM5A-KO NB4 cells) ± 3 kb are shown, sorted according to H3K4me2 binding intensity in control NB4. (B) ChIP-seq signal density plots showing the global levels of H3K4me3 and H3K4me2 at all genes ± 3 kb. (C) Box plot showing the expression levels of genes with increased H3K4me2 in KDM5A-KO, in the control and KDM5A-KO NB4 cells. Gene expression levels are presented as log2CPM. Statistical significance was determined by using Student’s t test. ****P < .0001.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Expressing, ChIP-sequencing, Binding Assay

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A KO led to the activation of PML-RARα target genes. (A) Volcano graph showing the gene expression changes upon KDM5A loss in NB4. (B) Box plot showing the expression of PML-RARα target genes in the control and KDM5A-KO NB4 cells. Gene expression levels are presented as log2CPM. The median log2CPM of control and KDM5A-KO NB4 cells were labeled. Statistical significance was determined by using Student’s t test. ****P < .0001 . (C) RT-qPCR examination of the expression of the select PML-RARα target genes. Statistical significance was determined by using Student’s t test. *P < .05, ***P < .001, ****P < .0001. (D) H3K4me2 ChIP-qPCR validation of the select PML-RARα target genes. The statistical significance was determined by using Student’s t test. **P < .01, ***P < .001, ****P < .0001. (E) Density plots of the indicated ChIP-seq signals over PML-RARα target genes ± 3 kb. (F) Heatmap showing the KDM5A binding in NB4 cells, and H3K4me2 and H3K4me3 levels in the control (Ctrl) and KDM5A-KO NB4 cells, over KDM5A bound PML-RARα target genes ± 3 kb, sorted according to KDM5A binding intensity. (G) ChIP-seq snapshots of 2 select PML-RARα target genes for KDM5A, H3K4me1, and input in NB4; H3K4me2, H3K4me3, and RNA-sequencing in the control and KDM5A-KO NB4 cells. TES, transcription end site; TSS, transcription start site.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Activation Assay, Expressing, Labeling, Quantitative RT-PCR, ChIP-sequencing, Binding Assay, RNA Sequencing Assay

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A inhibition induced NB4 differentiation. (A) Differentiation markers CD11b and CD11c in NB4 cells treated with indicated concentrations of a KDM5 inhibitor, CPI-455, measured by using flow cytometry. Statistical significance was determined by using Student’s t test. ***P < .001, ****P < .0001, P values for percentage of CD11b+ cells. #P < .05, ##P < .01, P values for percentage of CD11c+ cells. (B) The morphology of NB4 cells treated with dimethyl sulfoxide (DMSO) or 20 μM KDM5 inhibitor CPI-455. Bar = 15 μm. FACS, fluorescence-activated cell sorting.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Inhibition, Flow Cytometry, Fluorescence, FACS

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A KO lowered the exposure of ATRA to induce NB4 cell differentiation. (A) Proliferation assay of control (Ctrl) cells or KDM5A-KO cells treated with 0.5 nM ATRA or vehicle (dimethyl sulfoxide [DMSO]) and control cells treated with 10 nM ATRA for the indicated days. Viable cells were detected by using a trypan blue exclusion assay. The counts of the KO1 and KO2 cells with 0.5 nM ATRA were compared with the Ctrl cells with 10 nM ATRA. Statistical significance was determined by two-way analysis of variance. ****P < .0001. (B) CD11b+ or CD11c+ cells were detected by flow cytometry of control cells or KDM5A KO cells treated with indicated concentrations of ATRA for 4 days. ***P < .001, ###P < .001. (C) The morphology of control cells or KDM5A KO cells treated with 0.5 or 10 nM ATRA for 4 days. Bar = 15 μm. (D) The NBT test of control cells or KDM5A KO cells treated with indicated concentrations of ATRA or vehicle (DMSO) for 3 days. Values represent means ± standard deviation of the percentages of NBT-positive cells, n = 6.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Cell Differentiation, Proliferation Assay, Trypan Blue Exclusion Assay, Flow Cytometry, Standard Deviation

Journal: Blood Advances

Article Title: KDM5A suppresses PML-RARα target gene expression and APL differentiation through repressing H3K4me2

doi: 10.1182/bloodadvances.2020002819

Figure Lengend Snippet: KDM5A inhibition enhanced the differentiation-inducing effect of ATRA at a lower dose. (A)Proliferation assay of NB4 cells treated with 5 μM CPI-455 and/or 0.1 or 0.5 nM ATRA, 10 nM ATRA, or vehicle (dimethyl sulfoxide [DMSO]) for the indicated days. Viable cells were detected by using a trypan blue exclusion assay. Statistical significance was determined by two-way analysis of variance. ****P < .0001, ####P < .0001. (B) CD11b+ or CD11c+ cells were detected by flow cytometry of NB4 cells treated with 2.5 μM CPI-455 and/or 0.5 nM ATRA, or vehicle (DMSO) for 4 days. ***P < .001, *P < .05 (C) The morphology of NB4 cells treated with 2.5 μM CPI-455 and/or 0.5 nM ATRA, or vehicle (DMSO) for 4 days. Bar = 15 μm. (D) The NBT test of NB4 cells treated with 2.5 μM CPI-455 and/or 0.5 nM ATRA, or vehicle (DMSO) for 3 days. The values represent mean ± standard deviation of the percentages of NBT-positive cells, n = 6. FACS, fluorescence-activated cell sorting.

Article Snippet: Antibodies used for ChIP were as follows: H3K4me1 (Abcam, no. 8895), H3K4me2 (Abcam, no. 32356), H3K4me3 (Active Motif, no. 39159), KDM5A (Bethyl Laboratories, A300-897A), and anti–immunoglobulin G rabbit (Jackson ImmunoResearch Laboratories, lot no. 134230).

Techniques: Inhibition, Proliferation Assay, Trypan Blue Exclusion Assay, Flow Cytometry, Standard Deviation, Fluorescence, FACS