Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 4 Effect of ectopic expression of the junD gene on CDK4 transcription

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing

Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 5 Effect of ectopic expression of the junD gene on CDK4-promoter activity after deletion mutation of AP-1 binding site

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing, Activity Assay, Mutagenesis, Binding Assay

Journal: Biochemical Journal

Article Title: Induced JunD in intestinal epithelial cells represses CDK4 transcription through its proximal promoter region following polyamine depletion

doi: 10.1042/bj20061436

Figure Lengend Snippet: Figure 7 Effect of ectopic expression of the junD gene on CDK4-promoter luciferase reporter activity: AP-1 point mutation within CDK4-promoter

Article Snippet: The plasmid clone (pRSV-hjD) containing the human junD gene was obtained from ATCC.

Techniques: Expressing, Luciferase, Activity Assay, Mutagenesis

Journal: Cancer Cell

Article Title: Retinoic acid receptor activation reprograms senescence response and enhances anti-tumor activity of natural killer cells

doi: 10.1016/j.ccell.2024.02.004

Figure Lengend Snippet:

Article Snippet: c-JUN peptide , Tocris , Cat# 1989.

Techniques: Control, Purification, Recombinant, Staining, Reverse Transcription, Cell Isolation, Isolation, cDNA Synthesis, Gene Expression, shRNA, Plasmid Preparation, CRISPR, Software, Transfection, In Vitro, Imaging

Journal: Oncotarget

Article Title: Dual oxidase 2 and pancreatic adenocarcinoma: IFN-γ-mediated dual oxidase 2 overexpression results in H 2 O 2 -induced, ERK-associated up-regulation of HIF-1α and VEGF-A

doi: 10.18632/oncotarget.12032

Figure Lengend Snippet: ( A ) DUOX2 (left) and VEGF-A (right) expression in BxPC-3 and AsPC-1 cells with or without IFN-γ treatment for 24 h, as determined by quantitative RT-PCR. *** P < 0.001 vs. solvent-treated cells. ( B ) The same material as in A was analyzed for JunB (left) and c-Jun (right) expression. *** P < 0.001 vs. solvent-treated cells. ( C ) Western analysis of WCEs from BxPC-3 and AsPC-1 cells treated with IFN-γ for the indicated times. β-actin served as a loading control. The data are expressed as the mean ± SD of at least three independent experiments (A–B) or are representative of at least three independent experiments (C).

Article Snippet: Human β-actin (Hs99999903_m1), DUOX1 (Hs00213694_ml), DUOX2 (Hs00204187_m1), DUOXA2 (Hs01595310_g1), HIF-1a (Hs00153153_m1), c-Jun (Hs00277190_s1), JunB (Hs00357891_s1), Sp1 (Hs00916518_m1), Sp3 (Hs01595811_m1), and VEGF-A (Hs00900055_m1) primers were from Life Technologies (Carlsbad, CA, USA).

Techniques: Expressing, Quantitative RT-PCR, Solvent, Western Blot, Control

Journal: Oncotarget

Article Title: Dual oxidase 2 and pancreatic adenocarcinoma: IFN-γ-mediated dual oxidase 2 overexpression results in H 2 O 2 -induced, ERK-associated up-regulation of HIF-1α and VEGF-A

doi: 10.18632/oncotarget.12032

Figure Lengend Snippet: By engaging its cognate receptor (IFN-γR) on pancreatic cancer cells, IFN-γ up-regulates functional DUOX2/DUOXA2 expression that leads to enhanced H 2 O 2 production and oxidative inactivation of PTPs. PTP inactivation could then contribute to ERK, RSK, and S6 phosphorylation, leading to accumulation of HIF-1α and subsequent pro-angiogenic signal transduction, as well as concomitantly increasing expression of AP-1 (consisting of JunB/c-Jun). Binding of c-Jun and JunB to its transcriptional response element (ARE, AP-1 response element) in the nucleus, along with binding of the constitutively expressed transcription factor Sp1 to the GC box, appears to stimulate VEGF-A expression in BxPC-3 cells.

Article Snippet: Human β-actin (Hs99999903_m1), DUOX1 (Hs00213694_ml), DUOX2 (Hs00204187_m1), DUOXA2 (Hs01595310_g1), HIF-1a (Hs00153153_m1), c-Jun (Hs00277190_s1), JunB (Hs00357891_s1), Sp1 (Hs00916518_m1), Sp3 (Hs01595811_m1), and VEGF-A (Hs00900055_m1) primers were from Life Technologies (Carlsbad, CA, USA).

Techniques: Functional Assay, Expressing, Phospho-proteomics, Transduction, Binding Assay

Journal:

Article Title: Bile acids and cytokines inhibit the human cholesterol 7?-hydroxylase gene via the JNK/c-Jun pathway

doi: 10.1002/hep.21183

Figure Lengend Snippet: Effect of IL-1β and CDCA on mRNA expression in human primary hepatocytes. Human primary hepatocytes were treated with IL-1β (10 ng/ml) or CDCA (50 μM) for time periods indicated. Quantitative Real time PCR was performed as described in Materials and Methods. The relative mRNA levels of CYP7A1, HNF4α, SHP and c-Jun were calculated with respect to the UBC mRNA. The mRNA expression at 0 hr (un-treated) was set as 1. The error bars represent the standard deviation of from three different experiments. N=3.

Article Snippet: Assay-on-Demand PCR primers and Taqman MGB probe mix used are: human CYP7A1 (Cat. # Hs00167982_m1); human HNF4α(Cat. #: Hs00230853_m1); human cJun (Cat. #: Hs00277190_s1); human SHP (Cat. #: Hs00222677_m1); human CYP8B1 (Cat. #: Hs00244754_s1) and UBC (Cat. # Hs00824723_m1) (Applied Biosystems, Foster City, CA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal:

Article Title: Bile acids and cytokines inhibit the human cholesterol 7?-hydroxylase gene via the JNK/c-Jun pathway

doi: 10.1002/hep.21183

Figure Lengend Snippet: Co-immunoprecipitation assay of HNF4α and c-Jun interaction-HepG2 cell extracts treated with IL-1β (5 ng/ml) for the time indicated were immunoprecipitated with rabbit anti-HNF4α antibody as described in Materials and Methods. Immunobolot analysis was performed with goat anti-HNF4α, anti-c-Jun and anti-phospho c-Jun antibodies. Five percent of the cell lysate was immunoblotted with anti-actin antibody as internal control. Rabbit non-immune IgG was used as a negative control. The ratios of c-Jun to HNF4α and phospho-c-Jun to HNF4α are indicated below the panels.

Article Snippet: Assay-on-Demand PCR primers and Taqman MGB probe mix used are: human CYP7A1 (Cat. # Hs00167982_m1); human HNF4α(Cat. #: Hs00230853_m1); human cJun (Cat. #: Hs00277190_s1); human SHP (Cat. #: Hs00222677_m1); human CYP8B1 (Cat. #: Hs00244754_s1) and UBC (Cat. # Hs00824723_m1) (Applied Biosystems, Foster City, CA).

Techniques: Co-Immunoprecipitation Assay, Immunoprecipitation, Control, Negative Control

Journal:

Article Title: Bile acids and cytokines inhibit the human cholesterol 7?-hydroxylase gene via the JNK/c-Jun pathway

doi: 10.1002/hep.21183

Figure Lengend Snippet: Chromatin immunoprecipitation assay of HNF4α, PGC-1α, and c-Jun binding to the CYP7A1 chromatin. A. Anti-HNF4α antibody was used to precipitate chromatin from HepG2 cells treated with IL-1β (10 ng/ml) or CDCA (50 μM). B. Anti-c-Jun antibody was used to immunoprecipitate chromatin. C. HepG2 cells were transfected with cJun or JNK1 expression plasmid or pcDNA3 empty vector, and anti-HNF4α antibody was used to immunoprecipitate chromatin. D. HepG2 cells were transfected with HA-PGC-1α, and co-transfected with a c-Jun expression plasmid or pcDNA3 empty vector. A 391 bp fragment containing the BAREI and BAREII regions of the CYP7A1 promoter was PCR amplified and analyzed on a 1.5% agarose gel.

Article Snippet: Assay-on-Demand PCR primers and Taqman MGB probe mix used are: human CYP7A1 (Cat. # Hs00167982_m1); human HNF4α(Cat. #: Hs00230853_m1); human cJun (Cat. #: Hs00277190_s1); human SHP (Cat. #: Hs00222677_m1); human CYP8B1 (Cat. #: Hs00244754_s1) and UBC (Cat. # Hs00824723_m1) (Applied Biosystems, Foster City, CA).

Techniques: Chromatin Immunoprecipitation, Binding Assay, Transfection, Expressing, Plasmid Preparation, Amplification, Agarose Gel Electrophoresis

Journal:

Article Title: Bile acids and cytokines inhibit the human cholesterol 7?-hydroxylase gene via the JNK/c-Jun pathway

doi: 10.1002/hep.21183

Figure Lengend Snippet: Mechanisms of bile acid and cytokine inhibition of CYP7A1 gene transcription. Bile acids induce IL-1β synthesis and release from Kupffer cells. These cytokines activate JNK pathway in hepatocytes. The JNK pathway inhibits CYP7A1 via three possible mechanisms. 1) JNK inhibits HNF4α mRNA and protein expressions. 2) JNK phosphorylates HNF4α and reduces its DNA binding and trans-activation activity. 3) JNK induces c-Jun, which interacts with HNF4α and blocks HNF4α interaction with PGC-1α and results in inhibition of CYP7A1.

Article Snippet: Assay-on-Demand PCR primers and Taqman MGB probe mix used are: human CYP7A1 (Cat. # Hs00167982_m1); human HNF4α(Cat. #: Hs00230853_m1); human cJun (Cat. #: Hs00277190_s1); human SHP (Cat. #: Hs00222677_m1); human CYP8B1 (Cat. #: Hs00244754_s1) and UBC (Cat. # Hs00824723_m1) (Applied Biosystems, Foster City, CA).

Techniques: Inhibition, Binding Assay, Activation Assay, Activity Assay

Journal: Oncogene

Article Title: Suppression of Jab1/CSN5 induces radiation- and chemo-sensitivity in nasopharyngeal carcinoma through changes to the DNA damage and repair pathways

doi: 10.1038/onc.2012.294

Figure Lengend Snippet: Effect of Jab1 depletion on the sensitivity of NPC cells to cisplatin, IR and UV irradiation. (a) Jab1 expression in NPC cells. CNE1, CNE2, and HONE1 cells in the logarithmic growth phase were collected and lysed, followed by western blotting for Jab1. β-actin was used as a loading control. (Right) Quantification of Jab1 expression. (b, c, d, e) Effect of knockdown or ectopic overexpression of Jab1 on the sensitivity of NPC cells to cisplatin, IR and UV. HONE1 (b) and CNE2 (c) cells were transiently transfected with a Jab1 siRNA (Jab1-si) or scrambled control siRNA (Cont-si), CNE2 (d) cells were stably transfected with Jab1 shRNA (sh-Jab1) or control shRNA (sh-Cont), CNE1 (e) cells were transfected with ectopic Jab1 (Myc-Jab1) or a control vector (pcDNA). Cells were then analyzed for their Jab1 expression level (left) and response to cisplatin (CP), IR irradiation (middle) UV (right) as described in the Materials and Methods. Data represent three independent experiments, mean±s.d. * P <0.05, **P <0.01.

Article Snippet: TaqMan ® gene expression assays for the human Jab1/CSN5 (Hs00272789_m1), human Rad51 (Hs00153418_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Hs99999905_m1) were from Applied Biosystems/Life Technology (Carlsbad, CA, USA).

Techniques: Irradiation, Expressing, Western Blot, Over Expression, Transfection, Stable Transfection, shRNA, Plasmid Preparation

Journal: Oncogene

Article Title: Suppression of Jab1/CSN5 induces radiation- and chemo-sensitivity in nasopharyngeal carcinoma through changes to the DNA damage and repair pathways

doi: 10.1038/onc.2012.294

Figure Lengend Snippet: Effect of Jab1 on DNA damage-induced apoptosis. HONE1 cells were transiently transfected with either Jab1 siRNA (Jab1-si) or scrambled control siRNA (Cont-si) and treated with cisplatin (CP) (a) or IR (b) or UV (c) at the indicated doses. Apoptosis was analyzed by PI staining (left) or detection of cleaved PARP and caspase-3 by western blotting (right). (c) CNE1 cells were transfected with ectopic Myc-Jab1 plasmid and treated with cisplatin (CP) or UV for 48 h. Apoptosis was analyzed by Hoechst 33342 staining for condensed nuclei (left) and detection of cleaved PARP and caspase-3 by western blotting (right). Data represent three independent experiments, mean±s.d. *P <0.05, **P <0.01.

Article Snippet: TaqMan ® gene expression assays for the human Jab1/CSN5 (Hs00272789_m1), human Rad51 (Hs00153418_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Hs99999905_m1) were from Applied Biosystems/Life Technology (Carlsbad, CA, USA).

Techniques: Transfection, Staining, Western Blot, Plasmid Preparation

Journal: Oncogene

Article Title: Suppression of Jab1/CSN5 induces radiation- and chemo-sensitivity in nasopharyngeal carcinoma through changes to the DNA damage and repair pathways

doi: 10.1038/onc.2012.294

Figure Lengend Snippet: Jab1 associates with Rad51 and contributes to cisplatin (CP), IR and UV resistance. (a) NPC cells were transfected with Myc-Jab1 plasmid DNA or siRNA or transduced with Jab1 adenovirus in the presence or absence of doxycycline (Dox) for 48 h. Cell lysates were prepared 48 h after transfection or from cells stably expressing control (sh-Cont) or Jab1 shRNA (sh-Jab1) and western blotting was performed with the Jab1 and Rad51 antibodies. (Right) Correlation plot of Jab1 and Rad51 blot intensities obtained from control and treated cells. A fair linear correlation was obtained. (b) NPC cells were transfected with siRNA for 48 h, cells lysates were prepared and the levels of Jab1, p53 and Rad51 were determined by western blotting. (c) sh-Cont or sh-Jab1 stable CNE2 cells were transfected with siRNA for 48 h, then total protein and RNA was extracted and expression protein or RNA levels of the Jab1 and p53 and Rad51 were quantified by western blotting (left) or real time qRT-PCR (right). (d) CNE2 cells stably expressing sh-Jab1 were transfected with pcDNA or HA-Rad51 plasmid DNA. (Left) Western blot analyses demonstrated the effective knockdown and ectopic expression of Jab1. (Right and bottom) Colonies were stained with crystal violet 10 days after CP, IR and UV exposure. (e) (Top) CNE2 cells stably expressing sh-Cont or sh-Jab1 were exposed to UV or IR or CP. (Bottom) Cells stably expressing sh-Jab1 were transfected with pcDNA or HA-Rad51 plasmid DNA and then exposed to CP, IR and UV. Cleaved caspase-3 and γ̃H2AX were examined 48 h after exposure. All data represent three independent experiments, mean±s.d. *P <0.05, **P <0.01.

Article Snippet: TaqMan ® gene expression assays for the human Jab1/CSN5 (Hs00272789_m1), human Rad51 (Hs00153418_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Hs99999905_m1) were from Applied Biosystems/Life Technology (Carlsbad, CA, USA).

Techniques: Transfection, Plasmid Preparation, Transduction, Stable Transfection, Expressing, shRNA, Western Blot, Quantitative RT-PCR, Staining

Journal: Oncogene

Article Title: Suppression of Jab1/CSN5 induces radiation- and chemo-sensitivity in nasopharyngeal carcinoma through changes to the DNA damage and repair pathways

doi: 10.1038/onc.2012.294

Figure Lengend Snippet: Role of Jab1 in the response of NPC cells to cisplatin (CP), IR and UV. CNE1 and CNE2 cells were transiently transfected with Jab1 siRNA (Jab1-si) or scrambled control siRNA (Cont-si) and treated with CP, IR or UV for 48 h. Apoptosis was analyzed by Annexin-V and PI staining (a) , detection of cleaved PARP (C-PARP) and caspase-3 (C-Caspase 3) (b, c and d) . The percentage of CNE2 cells in the sub-G1 phase are shown (b and c, right) . Data represent three independent experiments, mean±s.d. *P <0.05, **P <0.01.

Article Snippet: TaqMan ® gene expression assays for the human Jab1/CSN5 (Hs00272789_m1), human Rad51 (Hs00153418_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Hs99999905_m1) were from Applied Biosystems/Life Technology (Carlsbad, CA, USA).

Techniques: Transfection, Staining

Journal: Oncogene

Article Title: Suppression of Jab1/CSN5 induces radiation- and chemo-sensitivity in nasopharyngeal carcinoma through changes to the DNA damage and repair pathways

doi: 10.1038/onc.2012.294

Figure Lengend Snippet: Knockdown of Jab1 impairs DNA repair and decreases expression of the DNA repair gene Rad51 . (a) (Left) RPPA analysis of cisplatin (CP)- and UV- treated NPC cells. Cell lysates were harvested after treatment with 5 μM CP or 40 J/m 2 UV for 48 h. Samples were organized by treatment, which is indicated on the right side of the heat-map. (Middle and Right) The response of the cells to CP or UV was normalized against the response of untreated cells. (c, d) HONE1 cells were transfected with Jab1 or control siRNA 48 h before CP, UV or IR exposure. Cell lysates were prepared 48 h after CP, UV or IR exposure and western blotting was performed with the indicated antibodies related to DNA damage and repair proteins.

Article Snippet: TaqMan ® gene expression assays for the human Jab1/CSN5 (Hs00272789_m1), human Rad51 (Hs00153418_m1), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Hs99999905_m1) were from Applied Biosystems/Life Technology (Carlsbad, CA, USA).

Techniques: Expressing, Transfection, Western Blot

Journal: JCI Insight

Article Title: Suppression of TCF4 promotes a ZC3H12A-mediated self-sustaining inflammatory feedback cycle involving IL-17RA/IL-17RE epidermal signaling

doi: 10.1172/jci.insight.172764

Figure Lengend Snippet: ( A ) Silencing TCF4 ( siRNA-mediated ) in N/TERT keratinocytes, but not NFKB1 , JUN , or CEBPG , increases IL17C mRNA expression, which is further increased in the presence of TNF-α stimulation (10 ng/mL; n = 3, mean ± SEM, 2-way ANOVA with post hoc Tukey test. * P < 0.01; ** P < 0.005, *** P < 0.001, **** P < 0.0001; dashed line and the diamond demarcate P < 0.05 between 2 indicated groups via Student’s t test . ( B ) ATAC-Seq of human KCs isolated from fresh tissue biopsies identifies TCF4 binding sites in open chromatin regions of IL17C and ZC3H12A promoters. ( C ) Representative images of healthy normal and of lesional and nonlesional Ps skin demonstrates decreases in TCF4 (nuclear localization) and increases in IL-17C and ZC3H12A staining (using IHC; stained protein appears brown in color). Insets represent higher-magnification image. Scale bar: 100 μm; 20 μm (insets). ( D ) IL17C expression negatively correlates with TCF4 and positively correlates with ZC3H12A in lesional Ps and AD skin.

Article Snippet: Human primers (Thermo Fisher Scientific) used in this study were: TCF4, Hs00162613_m1; IL-17C, Hs00171163_m1; JUN, Hs99999141_s1;CEBPG, Hs01922818_s1; IL1B, Hs01555410_m1; IL36G, Hs00219742_m1; CCL20, Hs00355476_m1; CXCL1, Hs00236937_m1; S100A7, Hs00161488_m1; DEFB4, Hs00175474_m1; IVL, Hs00846307_s1; FLG, Hs00856927_g1; LOR, Hs01894962_s1; SPRR2A, Hs03046643_s1; ZC3H12A, Hs00962356_m1; NFKBIZ, Hs00230071_m1; and RPLP0, Hs99999902_m1.

Techniques: Expressing, Isolation, Binding Assay, Staining