Journal: Epigenetics & Chromatin

Article Title: The pattern of histone H3 epigenetic posttranslational modifications is regulated by the VRK1 chromatin kinase

doi: 10.1186/s13072-023-00494-7

Figure Lengend Snippet: Inhibitors of histone PTMs

Article Snippet: KDM1A (LSD1) , pCMV6 , Myc-DDK , Mammalian , Origene, #RC208480.

Techniques: Concentration Assay

Journal: Epigenetics & Chromatin

Article Title: The pattern of histone H3 epigenetic posttranslational modifications is regulated by the VRK1 chromatin kinase

doi: 10.1186/s13072-023-00494-7

Figure Lengend Snippet: Plasmids

Article Snippet: KDM1A (LSD1) , pCMV6 , Myc-DDK , Mammalian , Origene, #RC208480.

Techniques: Plasmid Preparation, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: Staining patterns for LSD1 and SNAIL. LSD1 and SNAIL expressions were assessed using immunohistochemical staining. The staining intensities varied from ( a , e ) negative to ( b , f ) weak, ( c , g ) moderate, and ( d , h ) high positive nuclear staining for ( a – d ) LSD1 and ( e – h ) SNAIL.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Staining, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: LSD1 and SNAIL expression by tissue type and by primary tumor site. LSD1 and SNAIL expression were assessed in tissue types and primary tumor sites using the H-Score. ( a ) Primary tumors show a lower LSD1 expression compared to local recurrences and higher expression compared to LN-metastases. ( b ) Primary tumors that originate in the hypopharynx show a significantly higher LSD1 expression compared to laryngeal, oral, or oropharyngeal head and neck squamous cell carcinoma (HNSCC). ( c ) Primary tumors have a significantly lower SNAIL expression than local recurrences as well as LN- and distant metastases. ( d ) There were no significant differences in SNAIL expression between primary tumor locations. p -values were calculated using Mann–Whitney tests and are indicated: *- p < 0.05, **- p < 0.01, ***- p < 0.001, ****- p < 0.0001.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: LSD1 expression in primary HNSCC tumors by tumor site and patient and tumor characteristics. Shown are the average H-Score values of primary HNSCC separated by the site of origin and categorized by patient and tumor characteristics. Respective H-Score values of categories showing significant differences are bold. p -values were calculated using Mann–Whitney tests and are indicated: *- p < 0.05, **- p < 0.01, ***- p < 0.001.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: Association between LSD1 and SNAIL expression in primary HNSCC. High LSD1 expression is significantly associated with high SNAIL expression in primary HNSCC. The p -value was calculated using Pearsons’s Chi-squared test.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: Association between LSD1 and SNAIL expression considering all available tissue samples. LSD1 and SNAIL expression are significantly associated when all available tissue samples are considered. The p -value was calculated using Pearsons’s Chi-squared test.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: LSD1 and SNAIL staining in matched cores. LSD1 and SNAIL stained cores of identical samples showed similar staining intensities. ( a , b ) Samples were considered negative for both LSD1 and SNAIL. ( c , d ) Samples considered moderate or high positive for both LSD1 and SNAIL.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Staining

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: Kaplan–Meier analysis of primary HNSCC in general. Five-year survival curves showing the differences in ( a ) overall and ( b ) progression-free survival between low and high LSD1, SNAIL, or combined LSD1 and SNAIL expression in primary HNSCC. The corresponding p -values are shown in each panel and were calculated using two-sided log-rank testing.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Elevated LSD1 and SNAIL Expression Indicate Poor Prognosis in Hypopharynx Carcinoma

doi: 10.3390/ijms23095075

Figure Lengend Snippet: Kaplan–Meier survival analysis of primary hypopharyngeal HNSCC. Primary tumors originating in the hypopharynx were categorized into groups showing low and high LSD1, SNAIL, or combined LSD1 and SNAIL expression, and their differences in ( a ) overall and ( b ) progression-free survival were assessed using the Kaplan–Meier method. The corresponding p -values are shown in each panel and were calculated using two-sided log-rank testing.

Article Snippet: LSD1 antibody 1B2E5 (monoclonal, mouse, dilution 1:350, nb 100 1762; Novus Biological, Littleton, CO, USA) and the SNAIL antibody AF6032 (polyclonal, rabbit, dilution 1:1000, AF6032; Affinity Biosciences, Cincinnati, OH, USA) were used as primary antibodies.

Techniques: Expressing

Journal: Cancer Research

Article Title: Lysine-Specific Demethylase 1 Is Strongly Expressed in Poorly Differentiated Neuroblastoma: Implications for Therapy

doi: 10.1158/0008-5472.can-08-1735

Figure Lengend Snippet: Figure 1. A, immunohistochemical staining of LSD1 in neuroblastic tumors. In poorly differentiated neuroblastomas (I and II), nuclear LSD1 staining is observed in almost all tumor cells. In contrast, in benign ganglioneuroblastomas (III)/ganglioneuroma (IV), LSD1 staining is mild or absent. The few signals marked by arrows in III and IV represent nucleoli of differentiated ganglia. Infiltrating leukocytes (L) in II, and Schwannian stroma do not display any immunoreactivity for LSD1 (III/IV). B, a tissue microarray with 99 primary neuroblastic tumors was used to analyze LSD1 expression in neuroblastoma and its benign derivates. Expression was significantly higher in poorly differentiated neuroblastomas (NB pd) than in differentiated neuroblastomas (NB diff) or ganglioneuroblastomas/ganglioneuromas (GNB/GN). C, LSD1 mRNA expression in an independant cohort of 110 neuroblastic tumors analyzed with Affymetrix microarrays. D, Kaplan-Meier analysis of 110 neuroblastic tumors shows that low LSD1 mRNA expression levels are predictive of EFS.

Article Snippet: Immunoprecipitation was performed with specific antibodies to H3K4me2 (Abcam) and LSD1 (Novus Biologicals) on protein A coupled Dynabeads (Invitrogen).

Techniques: Immunohistochemical staining, Staining, Microarray, Expressing

Journal: Cancer Research

Article Title: Lysine-Specific Demethylase 1 Is Strongly Expressed in Poorly Differentiated Neuroblastoma: Implications for Therapy

doi: 10.1158/0008-5472.can-08-1735

Figure Lengend Snippet: Figure 2. A, LSD1 protein expression in different neuroblastoma cell lines. h-Actin was used as the loading control. B, treatment of SH-SY5Y cells with all-trans retinoic acid (RA) resulted in a significant increase in the number and length of neurites, which served as an indicator of differentiated phenotype. C, after 12 d, retinoic acid treatment induced differentiation of SH-SY5Y and BE(2)-C neuroblastoma cells. LSD1 protein levels were significantly reduced in both cell lines. h-Actin served as the loading control.

Article Snippet: Immunoprecipitation was performed with specific antibodies to H3K4me2 (Abcam) and LSD1 (Novus Biologicals) on protein A coupled Dynabeads (Invitrogen).

Techniques: Expressing, Control

Journal: Cancer Research

Article Title: Lysine-Specific Demethylase 1 Is Strongly Expressed in Poorly Differentiated Neuroblastoma: Implications for Therapy

doi: 10.1158/0008-5472.can-08-1735

Figure Lengend Snippet: Figure 3. A, transfection of SH-SY5Y neuroblastoma cells with LSD1-directed siRNA resulted in reproducible knockdown of LSD1 protein levels. h-Actin served as the loading control. B, SHEP and LAN1 cells were transfected with LSD1-directed siRNA. ChIP (left) was performed with the indicated antibodies. The precipitated DNA was amplified by PCR using primers flanking the TFPl2 proximal locus or XRCC5 proximal locus. siRNA-mediated knockdown of LSD1 was verified by quantitative PCR analysis (right). C, MTT assay of SH-SY5Y treated with siRNA against LSD1 detected a significant reduction in cell number after 72 h incubation (left). Phase-contrast microscopy of SH-SY5Y cells transfected either with siRNA against LSD1 or scrambled control siRNA. Phenotypic changes were observed 72 h posttreatment (right). D, microarray analysis of SH-SY5Y cells treated with siRNA directed against LSD1 or with scrambled control siRNA revealed an induction of genes involved in differentiation and neurite dynamics (red arrow). siRNA-mediated knockdown of LSD1/AOF2 resulted in reduction of LSD1 mRNA as expected (blue arrow).

Article Snippet: Immunoprecipitation was performed with specific antibodies to H3K4me2 (Abcam) and LSD1 (Novus Biologicals) on protein A coupled Dynabeads (Invitrogen).

Techniques: Transfection, Knockdown, Control, Amplification, Real-time Polymerase Chain Reaction, MTT Assay, Incubation, Microscopy, Microarray

Journal: Cancer Research

Article Title: Lysine-Specific Demethylase 1 Is Strongly Expressed in Poorly Differentiated Neuroblastoma: Implications for Therapy

doi: 10.1158/0008-5472.can-08-1735

Figure Lengend Snippet: Figure 4. A, treatment of SHEP (red), SH-SY5Y (green), or LAN-1 (blue) neuroblastoma cells with pargyline, tranylcypromine, or clorgyline resulted in extensive reduction of cell numbers and MTT uptake. B, Western blot analysis confirmed an accumulation of H3K4 dimethylation upon treatment with MAOIs. In contrast, LSD1 protein levels were not affected. h-Actin served as the loading control. C, relative tumor weight of SH-SY5Y xenografts in nude mice treated with 2 mg tranylcypromine (MAOI) or control (NaCl). Mice were sacrificed, and the tumors were weighed 21 d after s.c. tumor cell inoculation. Tumors in mice receiving tranylcypromine were significantly smaller than mice receiving saline only. D, histologic appearance of tumors treated without MAOI (saline, I) or with 2 mg tranylcypromine (II) for 21 d. Sections were stained with H&E. Note the massive necrosis and hemorrhage of tumors in the MAOI-treated xenografts (II).

Article Snippet: Immunoprecipitation was performed with specific antibodies to H3K4me2 (Abcam) and LSD1 (Novus Biologicals) on protein A coupled Dynabeads (Invitrogen).

Techniques: Western Blot, Control, Saline, Staining

Journal: The international journal of biochemistry & cell biology

Article Title: SNF5 promotes IL-1β expression via H3K4me1 in atherosclerosis induced by homocysteine.

doi: 10.1016/j.biocel.2021.105974

Figure Lengend Snippet: Fig. 3. KDM1A negatively regulates the expression of H3K4me1 in macrophage. A. H3K4 methylation of IL-1β gene in 7 cell lines were obtained from UCSC database. B. IL-1β is regulated by H3K4 methylation in human aorta and three kinds of macrophages. The data is derived from the Ensemble database. C. Western blot analysis of the H3K4 mono, di-and tri methylations in the macrophages treated with Hcy, and the H3 was used as loading control (n = 3). D. Western blot analysis of the H3K4me1 in ApoE−/−mice fed with normal diet and high methionine diet (n = 6). E. qRT-PCR was used to detect the mRNA expression of KDM1A and KDM1B in Hcy-treated macrophages (n = 3). F. The KDM1A and KDM1B protein expression in Hcy-treated macrophages (n = 3). G. The mRNA and protein expression of KDM1A in ApoE−/−mice fed with high methionine diet (n = 6). H. Photomicrographs of mouse aortic root immunostained with MOMA-2 (green), KDM1A (red), and DAPI (blue). Scale bar, 200 μm. I. The H3K4me1 in the macrophages after infection with LV-GFP, LV- KDM1A and treatment with Hcy (n = 3). All experiments were repeated at least 3 times, and the data are expressed as the Mean ± SD. **P < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

Article Snippet: Immunodetection was carried out using SNF5 antibody (ab222519, Abcam, UK), IL-1β antibody (ab9722, Abcam, UK), Histone H3(mono/di/tri methyl K4) antibody (ab8895/ab32356/ab8550, Abcam, UK), KDM1A antibody (NBP1− 76688, NOVUS, USA), KDM1B antibody (ab193080, Abcam, UK), and β-actin antibody (TA-09, ZSGB-Bio, China), following incubated with HRP-conjugated secondary antibodies (ZB-2301/ ZB-2305, ZSGB-Bio, China).

Techniques: Expressing, Methylation, Derivative Assay, Western Blot, Control, Quantitative RT-PCR, Infection

Journal: The international journal of biochemistry & cell biology

Article Title: SNF5 promotes IL-1β expression via H3K4me1 in atherosclerosis induced by homocysteine.

doi: 10.1016/j.biocel.2021.105974

Figure Lengend Snippet: Fig. 4. SNF5 promotes the expression of IL-1β by down-regulating the expression of KDM1A. A. H3K4me1 was detected by western blot after macrophages were transfected with LV-GFP and LV-SNF5 and treated with Hcy (n = 3). B. The H3K4me1 in the macrophages was detected by western blot after transfection with scramble, si-SNF5-1/2 and treatment with Hcy (n = 3). C. Chromatin immunoprecipitation (ChIP) analysis of the H3K4me1 levels at the IL-1β promoter after macrophages were infected with LV-GFP and LV-SNF5 under Hcy treatment. The ChIP-enriched DNA fragments of the IL-1β promoter using IgG and an anti- H3K4me1 antibody were amplified by PCR. Total input (5%) was used as a positive control (n = 3). D. ChIP analysis of the H3K4me1 levels at the IL-1β pro moter after macrophages were infected with scramble and si-SNF5-1/2 under Hcy treatment (n = 3). E. The KDM1A mRNA and protein expression levels in the macrophages after infection with LV-GFP, LV-SNF5 and treated with Hcy (n = 3). F. The mRNA and protein expression of KDM1A in the macrophages after transfected with scramble, si-SNF5-1/2 and treated with Hcy (n = 3). G. Immunofluorescence co-localization of SNF5 (red) and KDM1A (green) in macrophages. Representative photograph showed the co-localization of SNF5-KDM1A in merged images, nuclei were stained with DAPI (blue) (n = 6). Scale bar, 20 μm. H. Macrophages was treated with the Hcy and then total cell lysates were immunoprecipitated with SNF5 antibodies or control IgG, and proceeded for western blot analysis with the indicated antibodies. All experiments were repeated at least 3 times, and the data are expressed as the Mean ± SD. *P < 0.05, **P < 0.01. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article).

Article Snippet: Immunodetection was carried out using SNF5 antibody (ab222519, Abcam, UK), IL-1β antibody (ab9722, Abcam, UK), Histone H3(mono/di/tri methyl K4) antibody (ab8895/ab32356/ab8550, Abcam, UK), KDM1A antibody (NBP1− 76688, NOVUS, USA), KDM1B antibody (ab193080, Abcam, UK), and β-actin antibody (TA-09, ZSGB-Bio, China), following incubated with HRP-conjugated secondary antibodies (ZB-2301/ ZB-2305, ZSGB-Bio, China).

Techniques: Expressing, Western Blot, Transfection, Chromatin Immunoprecipitation, Infection, Amplification, Positive Control, Immunofluorescence, Staining, Immunoprecipitation, Control

Journal: Carcinogenesis

Article Title: Lysine-specific demethylase 1 (LSD1) is highly expressed in ER-negative breast cancers and a biomarker predicting aggressive biology.

doi: 10.1093/carcin/bgp324

Figure Lengend Snippet: Fig. 1. Overexpression of LSD1 in ER-negative breast tumors. (A) Immunohistochemical staining of LSD1 was observed in normal breast tissue and breast cancer (histological grades 2 and 3). (B) LSD1 expression level in 20 normal breast tissues samples, 26 ER-positive and 37 ER-negative breast carcinomas was analyzed with ELISA for LSD1. ER (þ), ER-positive; ER (), ER-negative. (C) LSD1 expression in normal and tumor tissue extracts was determined by western blot. Coomassie staining was used as the loading control. C, control: in vitro translated human LSD1; N, normal breast tissue; T, breast tumor tissue. (D) Statistical significance test of ELISA was done by two-sided non-parametrical Mann–Whitney U-test to analyze differences in expression levels among normal, ER-positive and ER-negative groups. LSD1 expression was significantly higher in ER-negative breast cancers than in ER-positive cancers or normal tissue (P , 0.001).

Article Snippet: The membranes were incubated for 1–2 h using the following antibodies and dilutions: a-LSD1 (Novus Biologicals) 1:1000; a-K4H3me2 (Abcam, Cambridge, UK) 1:1000; b-actin (Sigma-Aldrich, Hamburg, Germany) 1:5000.

Techniques: Over Expression, Immunohistochemical staining, Staining, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control, In Vitro, MANN-WHITNEY

Journal: Carcinogenesis

Article Title: Lysine-specific demethylase 1 (LSD1) is highly expressed in ER-negative breast cancers and a biomarker predicting aggressive biology.

doi: 10.1093/carcin/bgp324

Figure Lengend Snippet: Fig. 2. Reduction in cell growth and increase of global H3K4 methylation upon MAOIs treatment. (A) Four different breast cancer cells were treated with tranylcypromine and clorgyline for 72 h for MTT assay. MAOIs treatment resulted in extensive reduction of cell numbers. (B) Western blot analysis confirmed an accumulation of H3K4 dimethylation upon treatment with 10 or 30 lM tranylcypromine and 10 or 30 lM clorgyline for 24 h in all breast cancer lines. In contrast, LSD1 protein levels were not affected. b-actin served as the loading control. (C) Effect of estrogen treatment on the sensitivity to MAOIs was tested in T47D (ERþ) cells. Cells were treated with different concentrations of tranylcypromine in the absence or presence of 17b-estradiol (E2). Western blot analysis showed that E2 induced LSD1 protein. The unpaired t-tests were performed to show that the differences between two groups are significant (P , 0.05).

Article Snippet: The membranes were incubated for 1–2 h using the following antibodies and dilutions: a-LSD1 (Novus Biologicals) 1:1000; a-K4H3me2 (Abcam, Cambridge, UK) 1:1000; b-actin (Sigma-Aldrich, Hamburg, Germany) 1:5000.

Techniques: Methylation, MTT Assay, Western Blot, Control

Journal: Carcinogenesis

Article Title: Lysine-specific demethylase 1 (LSD1) is highly expressed in ER-negative breast cancers and a biomarker predicting aggressive biology.

doi: 10.1093/carcin/bgp324

Figure Lengend Snippet: Fig. 3. Decreased cellular growth upon siRNA-mediated knockdown of LSD1. (A) Knockdown of LSD1 protein levels was determined 6 days after transfection by western blot. b-Actin served as the loading control. A significant reduction in cell number was observed in MTT assay upon knockdown of LSD1 in T47D (B) for 6 days. Decreased cellular growth upon siRNA-mediated knockdown was observed in MDA-MB231 and MDA-MB 453 (C). MDA-MB231 and MDA-MB cells were treated with siRNA against LSD1 for 12 days.

Article Snippet: The membranes were incubated for 1–2 h using the following antibodies and dilutions: a-LSD1 (Novus Biologicals) 1:1000; a-K4H3me2 (Abcam, Cambridge, UK) 1:1000; b-actin (Sigma-Aldrich, Hamburg, Germany) 1:5000.

Techniques: Knockdown, Transfection, Western Blot, Control, MTT Assay

Journal: Carcinogenesis

Article Title: Lysine-specific demethylase 1 (LSD1) is highly expressed in ER-negative breast cancers and a biomarker predicting aggressive biology.

doi: 10.1093/carcin/bgp324

Figure Lengend Snippet: Fig. 5. Panels show ChIP/qPCR occupancy analysis of LSD1, H3K9me2 and H3K4me2 on p21, CCNA and ERBB2 genomic loci after treatment of cells with siRNA directed against LSD1 or with scrambled control siRNA. (A) LSD1 binds specific regions of the p21 promoter in MDA-MB 231 cells. The sites are located 370 bp (370) and 30 bp (30) upstream of the transcriptional start site. (B) Enrichment of H3K9 dimethylation in the proximal promoter region of CCNA2 or ERBB2 was observed upon knockdown of LSD1. In ChIP experiments, the sonicated chromatin of MDA-MB 231 and MCF7 cells was immunoprecipitated with a-LSD1, a-H3K9me2 and a-H3K4me2. The precipitated DNA was amplified by polymerase chain reaction using primers flanking the p21 (370), p21 (30), CCNA2 (70) or ERBB2 (250) genomic loci upstream of the transcriptional start site. Normalized values were calculated as ratio to histone H3. Significant differences are indicated with P , 0.05.

Article Snippet: The membranes were incubated for 1–2 h using the following antibodies and dilutions: a-LSD1 (Novus Biologicals) 1:1000; a-K4H3me2 (Abcam, Cambridge, UK) 1:1000; b-actin (Sigma-Aldrich, Hamburg, Germany) 1:5000.

Techniques: ChIP-qPCR, Control, Knockdown, Sonication, Immunoprecipitation, Polymerase Chain Reaction