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Suz12

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SUZ12 is a protein component of the Polycomb Repressive Complex 2 (PRC2), which is involved in the epigenetic regulation of gene expression. It plays a crucial role in the methylation of histone H3 at lysine 27, a modification associated with transcriptional repression. SUZ12 is essential for the catalytic activity and stability of the PRC2 complex.

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Lab products found in correlation

Trps1, by Santa Cruz Biotechnology (2 mentions) H3k27me3, by Merck Group (1 mentions) Histone h3, by Cell Signaling Technology (1 mentions) Ecl reagent, by GE Healthcare (1 mentions) β actin, by Merck Group (1 mentions) Gapdh sc 32233, by Santa Cruz Biotechnology (1 mentions) Fus tls sc 47711, by Santa Cruz Biotechnology (1 mentions) Image lab software, by Bio-Rad (1 mentions) Molecular imager chemidoc xrs, by Bio-Rad (1 mentions) H3k9me3, by Merck Group (1 mentions) H3k27me3, by Abcam (1 mentions) Sc 46264, by Santa Cruz Biotechnology (1 mentions) Kdm4b, by Abcam (1 mentions) H3k9me2, by Abcam (1 mentions) H3k9ac, by Abcam (1 mentions) Human anti ezh2 antibody, by Abcam (1 mentions) Ez magna rip kit, by Merck Group (1 mentions) Magna rip rna binding protein immunoprecipitation kit, by Merck Group (1 mentions) Dnmt1, by Santa Cruz Biotechnology (1 mentions) 5 azacytidine, by Merck Group (1 mentions)

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Anti-SUZ12 (4 citations)

10 protocols using suz12

1

Epigenetic Regulation Analysis

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Western blotting was performed with specific antibodies against Histone H3 (9715, Cell Signaling), β-Actin (A5441, Sigma), EZH2 (612667, BD), SUZ12 (46264, Santa Cruz), EED (28701, Santa Cruz), and H3K27me3 (07449, Millipore). Signals were detected by ECL reagents (GE Healthcare, Buckinghamshire, UK).
Nagarsheth N., Peng D., Kryczek I., Wu K., Li W., Zhao E., Zhao L., Wei S., Frankel T., Vatan L., Szeliga W., Dou Y., Owens S., Marquez V., Tao K., Huang E., Wang G, & Zou W. (2015). PRC2 epigenetically silences Th1-type chemokines to suppress effector T cell trafficking in colon cancer. Cancer research, 76(2), 275-282.
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2

Western Blot Analysis of N2a Cells

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Whole-cell protein extracts were prepared from N2a cells using RIPA buffer and subjected to western blot analysis performed using precasted NuPAGE SDS-PAGE gels and reagents from Invitrogen. The immunoblots were incubated with the following antibodies diluted in 5% skim milk in TBS-T: FUS/TLS (sc-47711, Santa Cruz, 1:2,000), GAPDH (sc-32233, Santa Cruz, 1:2,000), SUZ12 (SC-46264, Santa Cruz, 1:200), and TBP (sc-273, Santa Cruz, 1:200). All the images were captured using the Molecular Imager ChemiDoc XRS+ (Bio-Rad), and the densitometric analyses were performed using the associated Image Lab software (Bio-Rad). Full scan of western blots are presented in Supplementary Fig. 7.
Errichelli L., Dini Modigliani S., Laneve P., Colantoni A., Legnini I., Capauto D., Rosa A., De Santis R., Scarfò R., Peruzzi G., Lu L., Caffarelli E., Shneider N.A., Morlando M, & Bozzoni I. (2017). FUS affects circular RNA expression in murine embryonic stem cell-derived motor neurons. Nature Communications, 8, 14741.
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3

ChIP Protocol for Epigenetic Regulators

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ChIP experiments were performed as described previously [26 (link)]. Briefly, cells were cross-linked with 1% formaldehyde for 10 min at room temperature and cross-linked cells were lysed using lysis buffer and sonicated for 30 min. Then, Sonicated lysates were immunoprecipitated using antibodies with EZH2, DNMT1, LSD1, SUZ12 (Santa Cruz, USA) and IgG (Cell Signaling Technology, USA) antibodies.
Ba M.C., Long H., Cui S.Z., Gong Y.F., Yan Z.F., Wu Y.B, & Tu Y.N. (2017). Long noncoding RNA LINC00673 epigenetically suppresses KLF4 by interacting with EZH2 and DNMT1 in gastric cancer. Oncotarget, 8(56), 95542-95553.
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4

Nuclear Protein Extraction and Western Blot

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Nuclear extracts were prepared as described in 21 (link). Cells were washed twice with PBS1X and then resuspended and incubated for 15' in cold lysis buffer A plus inhibitors, followed by adding 10% Triton X100, cytoplasmic lysates were cleared by centrifugation. The pellet was resuspended and incubated for 15' in Nuclear Lysis Buffer (NLB), and then a spin down at high speed was necessary for recovering the nuclear proteins 21 (link). The proteins were quantitated and separated in SDS-polyacrylamide gels and transferred to nitrocellulose membranes, following overnight blocking with 10-20% non-fat milk at 4ºC. Membranes were incubated with the primary antibody at different dilutions at room temperature for 2 h and then incubated with a horseradish peroxidase-conjugated secondary antibody for 1 h at room temperature and detected by Chemiluminescence (Thermoscientific). Antibodies used were: HP1α (ab77256-abcam), H3K9me3 (07-442-millipore), H3K9me2 (ab1220-abcam), H3K9ac (ab4441-abcam), H3K27me3 (ab6002-abcam), KDM4A (PA5-25057-thermo scientific), KDM4B (ab27531-abcam), KDM4C (PA5-23065-thermo-scientific), KDM6A/UTX (ABE409-millipore), EZH2 (MABE362-millipore), SUZ12 (sc-46264-Santa Cruz) H3 (ab1791-abcam), Actin (JLA20-C-DSHB), Tubulin (E7-C- DSHB).
Gurrion C., Uriostegui M, & Zurita M. (2017). Heterochromatin Reduction Correlates with the Increase of the KDM4B and KDM6A Demethylases and the Expression of Pericentromeric DNA during the Acquisition of a Transformed Phenotype. Journal of Cancer, 8(14), 2866-2875.
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5

RIP Assay for EZH2, SUZ12, LSD1

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RIP experiments were performed using the EZ‐Magna RIP kit (Millipore, China) following the manufacturer's guidance. Human anti‐EZH2 antibody (Abcam, Shanghai, China), SUZ12 (Santa Cruz, Shanghai, China), LSD1 (Santa Cruz), or control IgG was used. The immunoprecipitated RNA was isolated and was further subjected to qRT‐PCR analysis.
Xiong Y., Kuang W., Lu S., Guo H., Wu M., Ye M, & Wu L. (2018). Long noncoding RNA HOXB13‐AS1 regulates HOXB13 gene methylation by interacting with EZH2 in glioma. Cancer Medicine, 7(9), 4718-4728.
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6

RNA Binding Protein Immunoprecipitation

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RNA immunoprecipitation (RIP) experiment was performed by using a Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, USA) following the manufacturer's instructions. MKN-45 and SGC-7901 cells were lysed and incubated with protein A Sepharose beads which were conjugated using antibodies with IgG, EZH2, DNMT1, LSD1 and SUZ12 (Santa Cruz, USA) at 4°C. The immunoprecipitated RNA was purified and detected by qRT-PCR.
Ba M.C., Long H., Cui S.Z., Gong Y.F., Yan Z.F., Wu Y.B, & Tu Y.N. (2017). Long noncoding RNA LINC00673 epigenetically suppresses KLF4 by interacting with EZH2 and DNMT1 in gastric cancer. Oncotarget, 8(56), 95542-95553.
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7

Epigenetic Regulation of Cellular Processes

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Suberoyl anilide hydroxamic acid (SAHA, a pan-HDAC inhibitor) and 3-Deazaneplanocin A (DZNep, a polycomb EZH2 subunit inhibitor) were purchased from Millipore, Temecula, CA. The DNA methylation inhibitor 5-AZA-Cytidine (AZA) was from Sigma, St. Louis, MO. Cells were treated with indicated concentration and incubated for 72 hours before harvesting. Agarose A/G plus is from Santa Cruz. The following antibodies were used as primary antibodies: SOX4 (Santa Cruz; 1:1000); EZH2 (BD; 1:4000); HDAC3 (Millipore; 1:4000); total histone H3 (Abcam; 1:5000); Tri-methylated histone H3 H3K27me3 (Millipore; 1:3000); SUZ12 (Santa Cruz; 1:1,000); EZH1 (Santa Cruz; 1:1,000); β-tubulin (Sigma-Aldrich; 1:5,000).
Koumangoye R.B., Andl T., Taubenslag K.J., Zilberman S.T., Taylor C.J., Loomans H.A, & Andl C.D. (2015). SOX4 interacts with EZH2 and HDAC3 to suppress microRNA-31 in invasive esophageal cancer cells. Molecular Cancer, 14, 24.
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8

Western Blot Analysis of Key Proteins

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Western blot analysis was performed as previously described [13] . The antibodies used in this study were: GAPDH (1:1000; cat no. ab9385, Abcam), TRPS1 (1:500; cat no. sc-26974, Santa Cruz), SUZ12 (1:500, cat no. sc-271325, Santa Cruz), E-cadherin (1:1000, cat no. #3195, CST), N-cadherin (1:1000; cat no. 18203, Abcam) and Vimentin (1:1000; cat no.#5741S, CST).
Hu J., Su P., Jiao M., Bai X., Qi M., Liu H., Wu Z., Sun J., Zhou G, & Han B. (2018). TRPS1 Suppresses Breast Cancer Epithelial-mesenchymal Transition Program as a Negative Regulator of SUZ12. Translational Oncology, 11(2), 416-425.
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9

ChIP Assay with Magnetic Beads

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ChIP was performed using the EZ-Magna ChIP assay kit (Millipore, Billerica, MD, USA) follow the manufacturer’s recommendation. Briefly, 1.0×107 cells were cross-linked with 1% formaldehyde. The cells were then sonicated, pre-cleaned, mixed with magnetic beads and incubated with 5-10 μg of antibody (TRPS1, cat no.sc-26974,Santa Cruz; SUZ12, cat no. sc-271325, Santa Cruz; H3K27 tri-methylation, cat no. 07-622, Millipore) per reaction overnight. After DNA purification, the enrichment of the DNA template was analyzed by PCR and RT-qPCR. Sequences of primers used were indicated in Table S1.
Hu J., Su P., Jiao M., Bai X., Qi M., Liu H., Wu Z., Sun J., Zhou G, & Han B. (2018). TRPS1 Suppresses Breast Cancer Epithelial-mesenchymal Transition Program as a Negative Regulator of SUZ12. Translational Oncology, 11(2), 416-425.
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10

Epigenetic Regulatory Protein Analysis

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Antibodies obtained from Cell Signaling Technology (Danvers, MA) included HDAC1 (5356), HDAC2 (2545), and HDAC3 (2632). β‐Actin (A‐2228) was obtained from Sigma‐Aldrich The antibody SUZ12 (catalog number: sc‐46264) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti‐EED (catalog number: 09‐774) and anti‐acetyl‐lysine (catalog number: 05‐515) were purchased from Millipore (Temecula, CA). The c‐Myc antibody (catalog number: 11667149001) was purchased from Roche Molecular Biochemicals (Indianapolis, IN).
Shi B., Behrens C., Vaghani V., Riquelme E.M., Rodriguez‐Canales J., Kadara H., Lin H., Lee J., Liu H., Wistuba I, & Simon G. (2019). Oncogenic enhancer of zeste homolog 2 is an actionable target in patients with non‐small cell lung cancer. Cancer Medicine, 8(14), 6383-6392.
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