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C maf

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C-Maf is a transcription factor that belongs to the Maf family. It plays a role in the regulation of gene expression.

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Lab products found in correlation

Matα1, by Abcam (3 mentions) C myc, by Abcam (3 mentions) β actin, by Abcam (3 mentions) Rnaimax, by Thermo Fisher Scientific (3 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) α 32p dctp, by PerkinElmer (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions) Bile acid, by Merck Group (1 mentions) C myc, by Thermo Fisher Scientific (1 mentions) Interleukin 6 il 6, by Merck Group (1 mentions) Seq chip, by Abcam (1 mentions) 5 aza 2 deoxycytidine, by Merck Group (1 mentions) γ 32p atp, by PerkinElmer (1 mentions) Enhanced chemiluminescence system, by GE Healthcare (1 mentions) Goat anti rabbit igg hrp antibody, by Santa Cruz Biotechnology (1 mentions) P stat4, by Abcam (1 mentions) P stat1, by Abcam (1 mentions) Stat4, by Abcam (1 mentions) T bet, by Abcam (1 mentions)

5 protocols using c maf

1

Western Blot and ChIP-Seq Workflow

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α-32P dCTP (6,000 Ci/mmol) was purchased from PerkinElmer (Boston, MA). Antibodies used for Western, ChIP, Seq-ChIP, and immunohistochemistry (IHC) to PHB1, MATα1, c-MYC, MAFG, c-MAF, β-ACTIN, and IgG were purchased from Abcam (Cambridge, MA). Lipofectamine 2000 and RNAi-Max were purchased from ThermoFisher (Carlsbad, CA). siRNAs to PHB1 (Cat# 4392422), c-MYC (5′-CGAUUCCUUCUAACAGAAtt-3′), MAFG (5′-CGGACUAGAGAGAGUUGCGtt-3′), c-MAF (5′-GCAUCGUGUACUUACCAGUtt-3′) and MAT1A (5′-GCACAACGAAGACAUCACGtt-3′) were purchased from ThermoFisher. Bile acids and interleukin-6 (IL-6) were from Sigma (St. Louis, MS).
Fan W., Yang H., Liu T., Wang J., Li T.W., Mavila N., Tang Y., Yang J., Peng H., Tu J., Annamalai A., Noureddin M., Krishnan A., Gores G.J., Martínez-Chantar M., Mato J.M, & Lu S.C. (2017). Prohibitin 1 Suppresses Liver Cancers Tumorigenesis in Mice and Human Hepatocellular and Cholangiocarcinoma cells. Hepatology (Baltimore, Md.), 65(4), 1249-1266.
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2

Investigating Transcriptional Regulators

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γ-32P ATP (3,000 Ci/mmol) and α-32P dCTP (6,000 Ci/mmol) were purchased from PerkinElmer (Boston, MA). Antibodies used for either Western and/or immunohistochemistry to MATα1, c-Myc, MafG, c-Maf, β-actin and IgG were purchased from Abcam (Cambridge, MA). Lipofectamine 2000 and RNAi-max were purchased from Invitrogen (Carlsbad, CA). siRNAs to c-Myc (5’-CGAUUCCUUCUAACAGAAtt-3’), MafG (5’-CGGACUAGAGAGAGUUGCGtt-3’), c-Maf (5’-GCAUCGUGUACUUACCAGUtt-3’) and MAT1A (5’-GCACAACGAAGACAUCACGtt-3’) were purchased from Ambion (Grand Island, NY). DNA methylation inhibitor 5-aza-2’-deoxycytidine was purchased from Sigma-Aldrich (St. Louis, MO). 3-Deazaneplanocin A hydrochloride was purchased from TOCRIS (Bristol, UK).
Yang H., Liu T., Wang J., Li T.W., Fan W., Peng H., Krishnan A., Gores G.J., Mato J.M, & Lu S.C. (2016). Deregulated Methionine Adenosyltransferase α1, c-Myc and Maf Proteins Interplay Promotes Cholangiocarcinoma Growth in Mice and Humans. Hepatology (Baltimore, Md.), 64(2), 439-455.
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3

Western Blot Analysis of Transcription Factors

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Total protein was isolated from PBMCs by using RIPA buffer (Abcam, Cambridge, United Kingdom). The amount of proteins was quantified by BCA Protein assay kit (Thermo Fisher Scientific, Rockford, IL, United States), with bovine serum albumin (BSA) as a standard. Each protein sample was loaded onto 10% SDS-polyacrylamide gel and transferred to nitrocellulose membrane (Schleicher & Schuell BioScience, Germany) for 90 min at 4°C, and blocked with 5% skim milk in TBST (1 M Tris–HCl, 5 M NaCl, 10% Tween-20) for 1 h at room temperature. The blots were incubated with anti-GATA3, -T-bet, -C-maf, -STAT1, -P-STAT1, -STAT4, -P-STAT4 or -beta-actin (all from Abcam) antibodies overnight at 4°C. Primary antibody-bound membranes were incubated with goat anti-rabbit IgG-HRP antibody (Santa Cruz Biotechnology, United States) for 1 h at room temperature. The target protein was visualized with enhanced chemiluminescence system (GE Healthcare Life Sciences, United States), followed by analysis using ChemiDoc XRS (Bio-Rad).
Kim H.W., Hong R., Choi E.Y., Yu K., Kim N., Hyeon J.Y., Cho K.K., Choi I.S, & Yun C.H. (2018). A Probiotic Mixture Regulates T Cell Balance and Reduces Atopic Dermatitis Symptoms in Mice. Frontiers in Microbiology, 9, 2414.
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4

Western Blot Analysis of Cell Signaling

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Cells were collected and lysed in RIPA lysis buffer (Yeasen, catalog #20101ES60) containing a protease inhibitor cocktail (Yeasen, catalog #20104ES08). Protein concentration was quantified by bicinchoninic acid assay (Thermal Fisher). Equal amounts of soluble protein were loaded and separated on a 10% SDS-PAGE, followed by transfer to nitrocellulose and then immunoblotting using primary antibodies, including p53 (CST, catalog #32532), p21 (abcam, catalog #ab109199), c-Myc (CST, catalog #13987 T), c-Maf (abcam, catalog #ab77071), p-STAT3 (CST, catalog #9145), t-STAT3 (CST, catalog #9139), PD-L1 (R&D, catalog #AF1019), Caspase 3 (CST, catalog #9665S), ZAP70 (CST, catalog #3165S), MDM2 (BD, catalog #556353), and β-actin (CST, catalog #3700S). HRP-conjugated secondary antibodies (Yeasen, catalog #33101ES60, catalog #33201ES60) were used at 1:5000 dilution.
Fang D.D., Tang Q., Kong Y., Wang Q., Gu J., Fang X., Zou P., Rong T., Wang J., Yang D, & Zhai Y. (2019). MDM2 inhibitor APG-115 synergizes with PD-1 blockade through enhancing antitumor immunity in the tumor microenvironment. Journal for Immunotherapy of Cancer, 7, 327.
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5

Immunohistochemical Analysis of Liver and CCA

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Formalin-fixed liver and CCA tissues embedded in paraffin were cut and stained with hematoxylin and eosin (H&E) for routine histology. Immunohistochemical staining of Matα1, c-Myc, MafG, c-Maf and IgG (Abcam, Cambridge, MA) was performed with Dako kit (Carpinteria, CA) or Abcam according to the manufacturer’s method. For quantifying immunohistochemical staining, a total of five fields at 100× magnification were randomly selected (minimum of 1000 cells total) and positive nuclei or cells were counted and expressed as a percentage of the total using the MetaMorph imaging software (Woburn, MA). Control with normal mouse IgG showed no staining.
Yang H., Liu T., Wang J., Li T.W., Fan W., Peng H., Krishnan A., Gores G.J., Mato J.M, & Lu S.C. (2016). Deregulated Methionine Adenosyltransferase α1, c-Myc and Maf Proteins Interplay Promotes Cholangiocarcinoma Growth in Mice and Humans. Hepatology (Baltimore, Md.), 64(2), 439-455.
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