Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: KDM4C expression in post-surgical, recurrent primary and metastatic sites, comparing to primary sites of non-recurrent CRC patients. a Expression levels of KDM4C mRNA in 124 CRC tissues and matched metastatic sites were analyzed by real-time PCR. The significant differences between primary tumor I (without paired metastatic tissues) and primary tumor II (with paired metastatic tissues, Metastasis II) were analyzed using the Wilcoxon signed-rank test. b Kaplan-Meier analyses of the correlations between KDM4C mRNA expression levels and overall survival (OS) of 124 CRC patients, and the median expression level was used as the cutoff. c The co-expression between KDM4C and MALAT1was showed in 124 cases from our datasets. d The mRNA expression of JMJD2C in 367 CRC primary tissues from TCGA dataset, and its association with CRC prognosis, including OS. e Immunohistochemical analysis of JMJD2C protein (encoded by KDM4C) in representative CRC and metastatic lung/liver tissues, including primary CRC tumor without paired metastatic tissues, and primary CRC tumor with paired metastatic tissues (scale bars, 100 μm, respectively). *, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Immunohistochemical staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: Translocation of JMJD2C protein from the cytoplasm into the nuclei in CRC cells in vitro. a - b Immunofluorescence detection of JMJD2C protein in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. c - d Western blot and quantitative assay of JMJD2C protein (nuclear and whole cell lysates) in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. *, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Translocation Assay, In Vitro, Immunofluorescence, Transfection, shRNA, Plasmid Preparation, Over Expression, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: JMJD2C promoted MALAT1 expression by regulating the histone methylation level of MALAT1 promoter. a Schematic diagram of the ChIP (Chromatin Immunoprecipitation) procedure for detecting the effect of JMJD2C on the histone methylation level of MALAT1 promoter in sites of H3K9m3 and H3K36m3. Purple ovals: no-targeting protein; Yellow ovals: targeting protein A; Green ovals: protein binding to targeting protein A. b JMJD2C, H3K9, H3K36 antibody and MALAT1 special primers were used to investigate the interaction between JMJD2C and MALAT1 promoter, IgG was used as the negative control. c-d MALAT1 promoter activities assay in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. e-f Real time PCR assay of MALAT1 levels in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. *, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Expressing, Methylation, Chromatin Immunoprecipitation, Protein Binding, Negative Control, Transfection, shRNA, Plasmid Preparation, Over Expression, Real-time Polymerase Chain Reaction

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: JMJD2C activated the β-catenin signaling pathway in CRC cells. a - b Immunofluorescence detection of β-catenin protein in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. c - d Western blot and quantitative assay of β-catenin protein (nuclear, cytoplasm and whole cell lysates) and β-catenin signaling downstream targets including c-Myc and ITGBL1 in HCT116 or LoVo cells transiently transfected with shRNA/NT vector, shRNA/JMJD2C vector, empty overexpression vector, or JMJD2C overexpression vector. *, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Immunofluorescence, Transfection, shRNA, Plasmid Preparation, Over Expression, Western Blot

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: JMJD2C promoted the metastasis of CRC cells in vivo. a-b HCT116-shRNA/NT, HCT116-shRNA/JMJD2C, HCT116-empty vector, and HCT116-JMJD2C +/+ cells were respectively injected into the tail vein of nude mice ( n = 6). After 42 days, the lung metastases images were observed and quantified by LB983 NIGHTOWL II system. c Quantification of lung metastatic nodules from 6 mice subjected to the indicated treatments. d-e Immunohistochemical and quantitative analysis of JMJD2C proteins on consecutive tissue microarray slides of lung metastatic nodules from 6 mice subjected to the indicated treatments. *, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: In Vivo, shRNA, Plasmid Preparation, Injection, Immunohistochemical staining, Microarray

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: JMJD2C elevated the expression of MALAT1 and β-catenin signaling related proteins in CRC lung metastasis mice models. a Real-time PCR was performed to detect the expression of MALAT1 in lung metastatic nodules from 6 mice subjected to the indicated treatments. b-c Western blot and quantitative assay of β-catenin protein (nuclear, cytoplasm and whole cell lysates) in the lung metastatic tissues from 6 mice subjected to the indicated treatments. d-e Immunohistochemical and quantitative analysis of ITGBL1 and c-Myc proteins on consecutive tissue microarray slides of lung metastatic nodules from 6 mice subjected to the indicated treatments.*, P < 0.05; **, P < 0.01 ( t test)

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining, Microarray

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: JMJD2C promotes colorectal cancer metastasis via regulating histone methylation of MALAT1 promoter and enhancing β-catenin signaling pathway

doi: 10.1186/s13046-019-1439-x

Figure Lengend Snippet: A schematic model of JMJD2C promoted CRC metastasis by regulating the histone methylation level of the MALAT1 promoter and enhancing β-catenin signaling pathway

Article Snippet: The primary antibodies used were JMJD2C (Santa Cruz, USA), β-catenin (CST, USA), c-Myc (CST, USA), ITGBL1 (Abcam, USA), PCNA (CST, USA) and GAPDH (CST, USA).

Techniques: Methylation

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: Embryo survival and kidney phenotypes of zebrafish embryos (96 untreated embryos and 96 embryos treated with kdm4c-MO injection). ( A ). Comparison of embryo survival rate between embryos treated with a single injection of 0.2 mM kdm4c-MO and untreated embryos (n = 96 in each group). ( B ). Pronephric phenotypes of zebrafish embryos after kdm4c-MO injection. Each kidney photo was taken from the dorsal view and at the developmental stage of 48 hpf of bTg(wt1b:EGFP) embryos. Compared with the untreated group, after kdm4c-MO injection, the renal tubules were tortuously dilated and the glomeruli were swollen. ( C ). Comparison of the defect rates of morphological phenotypes (tortuously dilated renal tubules and swollen glomeruli) of zebrafish embryos between untreated group and kdm4c-MO injection group. ( D ). Results of Western blotting for Kdm4c with zebrafish embryo kidneys with or without kdm4c-MO injection. Abbreviation: KDM4C, Lysine demethylase 4C; MO, morpholino oligonucleotides.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: Injection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: KDM4C inhibition increased cell apoptosis during oxidative stress in vitro. HEK293 cells treated with the KDM4C inhibitor, JIB04 (1 μM) were used for the study. H 2 O 2 (100 μM) treatment was used to induce oxidative stress. Cell apoptosis was analyzed by flow cytometry at the time points indicated in the figure. The percentages of dead cells at different time points after treatment are shown. Abbreviation: KDM4C, Lysine demethylase 4C.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: Inhibition, In Vitro, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: KDM4C depletion impaired autophagy during serum starvation in vitro. HEK293 cells transfected with KDM4C -sh plasmid were used for the study. Cells were harvested after serum starvation at the time points indicated in the figure. ( A ). Results of Western blotting for autophagy markers, LC3B, BCL1, and BCL2. ( B ). Results of immunofluorescence staining for LC3B in control and KDM4C knockdown HEK293 cells with or without serum starvation. ( C ). Relative ratio of fluorescence intensities of GFP-LC3B puncta with WT as the denominator. ( D ). Results of Western blotting for KDM4C in control and KDM4C knockdown cells. Abbreviation: K/D, knock down; KDM4C, Lysine demethylase 4C; WT, wild-type.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: In Vitro, Transfection, Plasmid Preparation, Western Blot, Immunofluorescence, Staining, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: Temporal changes in KDM4C and progression of kidney fibrosis during kidney injury. Wild-type mice receiving kidney ischemia-reperfusion injury (IRI) were used for this study. Kidney tissues, after IRI for the durations indicated in the figure, were harvested ( n = 8 for each group). ( A ). Western blotting results for KDM4C, H3K9M3, and HIF-1α. The average relative ratios of KDM4C and HIF-1α along the course of injury are shown. ( B ). Masson trichrome staining results of kidney tissues after IRI. Mild glomerulous and tubulointerstitial fibrosis, 3 days after receiving IRI-AKI. The severity of glomerulous and tubulointerstitial fibrosis was even more pronounced 2 weeks after IRI-AKI. Abbreviation: HIF, hypoxia-inducible factor; KDM4C, Lysine demethylase 4C.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: Western Blot, Staining

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: KDM4C depletion worsened acute kidney injury. Study animals receiving kidney ischemia-reperfusion injury were used for this study. Kidney tissues and serum after ischemia-reperfusion injury for the durations indicated in the figure were harvested ( n = 8 for each group). ( A ). Serum BUN and creatinine levels of wild-type and Kdm4c −/− mice after kidney ischemia-reperfusion injury. ( B ). Masson trichrome staining results of kidney tissues of wild-type and Kdm4c −/− mice after ischemia-reperfusion injury. ( C ). Representative results of kidney inflammatory cytokines of the study animals, 1 week after ischemia-reperfusion injury. ( D ). Results of Western blotting for KDM4C in the kidneys of wild-type and Kdm4c −/− mice at the age of 10 weeks. Abbreviation: BUN, blood urea nitrogen; CCL12, C-C motif ligand 12; CXCL1, C-X-C motif ligand 1; FGF, fibroblast growth factor; IGFBP-2, insulin-like growth factor binding protein-2; IL-1R, interleukin-1 receptor; KDM4C, lysine demethylase 4C. KIM-1, kidney injury molecule-1; NGAL, neutrophil gelatinase-associated lipocalin; WT, wide-type. * p < 0.05.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: Staining, Western Blot, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Essential Roles of the Histone Demethylase KDM4C in Renal Development and Acute Kidney Injury

doi: 10.3390/ijms23169318

Figure Lengend Snippet: Differential kidney expressions of genes between the wild-type and Kdm4c −/− mice. Kidney tissues of wild-type and Kdm4c −/− mice at the age of 10 weeks were used for this study ( n = 8 in each group). ( A ). Volcano plot showing differential kidney gene expressions between the wild-type and Kdm4c −/− mice. The results showed that 42 genes had higher expression levels, while 46 genes had lower expression levels in the Kdm4c −/− mice than in the wild-type mice ( p -value < 0.01, q -value < 0.01). ( B ). The KEGG pathway analysis results showed that Kdm4c deletion was also associated with a variety of neurodegenerative diseases and metabolic diseases. ( C ). GO enrichment analysis showed that the Kdm4c −/− mice had significantly different expressions in several pathways, including inner mitochondrial membrane protein complex and mitochondrial protein complex. ( D ). GSEA revealed that Kdm4c deletion was also associated with several pathways involved in mitochondrial function, including mitochondrial electron transport chain, oxidative phosphorylation, and mitochondrial membrane phospholipid-based signal pathways. Abbreviation: KDM4C, lysine demethylase 4C; KEGG, Kyoto Encyclopedia of Genes and Genomes; GSEA, gene set enrichment analysis; WT, wild-type.

Article Snippet: KDM4C knockdown was carried out using KDM4C human siRNA oligo duplex (Locus ID 23081) (Origene, Rockville, MD, USA).

Techniques: Expressing