Journal: BMC Nephrology

Article Title: Regulon analysis identifies protective FXR and CREB5 in proximal tubules in early diabetic kidney disease

doi: 10.1186/s12882-023-03239-6

Figure Lengend Snippet: Representative images of FXR ( A ) and CREB5 ( B ) immunostaining (green) in the cortex region of the kidney from DKD and MCD patients. The white arrows denote the nuclear localization of FXR or CREB5

Article Snippet: The antibodies used for western blotting included FXR (proteintech, 25055-1-AP), CREB5 (Bioss, bs-14053R), cleaved caspase-3 (Cell signaling, 05/2016), cleaved PARP1 (Proteintech, 66520-1-Ig), fibronectin (Proteintech, 66042-1-Ig), vimentin (Proteintech,10366-1-AP), E-cadherin (Proteintech, 20874-1-AP), GAPDH (Proteintech, 60004-1-Ig), and beta-tubulin (Bioworld, AP0064).

Techniques: Immunostaining

Journal: BMC Nephrology

Article Title: Regulon analysis identifies protective FXR and CREB5 in proximal tubules in early diabetic kidney disease

doi: 10.1186/s12882-023-03239-6

Figure Lengend Snippet: Knockdown of CREB5 sensitized HK2 cells to AGE-induced apoptosis. A Immunoblotting showed that siCREB5 significantly reduced CREB5 protein in HK2 cells. B CREB5 knockdown markedly enhanced the AGE-induced activation of caspase-3 and PARP1. All the results represent the data from three independent experiments. * p < 0.05; ** p < 0.01; # p < 0.05, statistical significance

Article Snippet: The antibodies used for western blotting included FXR (proteintech, 25055-1-AP), CREB5 (Bioss, bs-14053R), cleaved caspase-3 (Cell signaling, 05/2016), cleaved PARP1 (Proteintech, 66520-1-Ig), fibronectin (Proteintech, 66042-1-Ig), vimentin (Proteintech,10366-1-AP), E-cadherin (Proteintech, 20874-1-AP), GAPDH (Proteintech, 60004-1-Ig), and beta-tubulin (Bioworld, AP0064).

Techniques: Knockdown, Western Blot, Activation Assay

Journal: BMC Nephrology

Article Title: Regulon analysis identifies protective FXR and CREB5 in proximal tubules in early diabetic kidney disease

doi: 10.1186/s12882-023-03239-6

Figure Lengend Snippet: CREB5 and FXR knockdown aggravated AGE-induced EMT in HK2 cells. Immunoblotting of fibronectin, vimentin, E-cadherin in HK2 cells treated with siCREB5 ( A ) and siFXR ( B ) for 24 h, followed by treatment with 200 µg/ml AGE for 24 h. The results represent the data from three independent experiments. * p < 0.05; ** p < 0.01; *** p < 0.001; # p < 0.05; ### p < 0.001. statistically significant

Article Snippet: The antibodies used for western blotting included FXR (proteintech, 25055-1-AP), CREB5 (Bioss, bs-14053R), cleaved caspase-3 (Cell signaling, 05/2016), cleaved PARP1 (Proteintech, 66520-1-Ig), fibronectin (Proteintech, 66042-1-Ig), vimentin (Proteintech,10366-1-AP), E-cadherin (Proteintech, 20874-1-AP), GAPDH (Proteintech, 60004-1-Ig), and beta-tubulin (Bioworld, AP0064).

Techniques: Knockdown, Western Blot

Journal: The Journal of Clinical Endocrinology and Metabolism

Article Title: Characterizing 24-Hour Skeletal Muscle Gene Expression Alongside Metabolic and Endocrine Responses Under Diurnal Conditions

doi: 10.1210/clinem/dgae350

Figure Lengend Snippet: Gene expression assay targets in human skeletal muscle ( vastus lateralis )

Article Snippet: CREB5 , cAMP responsive element binding protein 5 , Hs00191719_m1.

Techniques: Gene Expression, Binding Assay

Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group

Article Title: Hyperthermia promotes exosome secretion by regulating Rab7b while increasing drug sensitivity in adriamycin-resistant breast cancer.

doi: 10.1080/02656736.2022.2029585

Figure Lengend Snippet: Figure 4. The validation experiment of hub genes. (A) The differences in gene expression of 10 hub genes were validated with RT-qPCR. (B) Western blot analysis of FOS, CREB5, MAPK8 and NFKB1 protein level. (C) The efficiency of siRNA to knockdown the expression of FOS and CREB5. (D) Cell proliferation in the si-FOS and si-CREB5 group was faster compared with that in the control group, using the CCK-8 assay. (E). Half-inhibition rate of adriamycin in MCF-7/ADR cells treated with si-FOS and si-CREB5. (F) Different expression of FOS and CERB5 between invasive breast cancer and normal breast tissue by TCGA database. Error bars represented the mean ± SD of at least three independent experiments, p < .05, p < .01, p < .001.

Article Snippet: The membrane was then probed with primary antibodies at 4 C overnight and with species-specific secondary antibodies at room temperature for 1 h. The primary antibodies, including FOS antibody (1:1000, Proteintech, 66590-1-Ig), NF-jB antibody (1:1000, Proteintech, 14220-1-AP), MAPK8 antibody (1:1000, Proteintech, 66210-1-Ig), CREB5 antibody (1:1000, Proteintech, 14196-1-AP), GAPDH antibody (1:5000, Proteintech, 60004-1-Ig), CD9 antibody (1:1000, Abcam, ab223052), CD63 antibody (1:1000, Abcam, ab68418), TSG101 Figure 1.

Techniques: Biomarker Discovery, Gene Expression, Quantitative RT-PCR, Western Blot, Knockdown, Expressing, Control, CCK-8 Assay, Inhibition

Journal: International journal of hyperthermia : the official journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group

Article Title: Hyperthermia promotes exosome secretion by regulating Rab7b while increasing drug sensitivity in adriamycin-resistant breast cancer.

doi: 10.1080/02656736.2022.2029585

Figure Lengend Snippet: Figure 5. Hyperthermia promotes the secretion of exosome in MCF-7/ADR cells. (A) The expression level of FOS and CREB5 in MCF-7/ADR exosomes after hyper- thermia. (B) Half-inhibition rate of adriamycin in MCF-7/ADR cells incubated with the exosomes which produced by MCF-7/ADR cells after hyperthermia. (C) The expression level of FOS and CREB5 in MCF-7/ADR after incubated with hyperthermia ADR/exo. (D) Particles concentration of exosomes derived from breast cancer cells analyzed by NTA. (E) Total protein amounts of exosomes derived from breast cancer cells analyzed by bicinchoninic acid (BCA) protein kit. (F) Transmission electron micrographs (TEM) of cells before or 6 h after hyperthermia. Intense extracellular vesicle shedding occurred 6 h after hyperthermia. Scale bar ¼1 lm. (G) Confocal microscope analysis of exosome uptake by MCF-7/ADR cells with or without hyperthermia treatment. Scale bar ¼20 lm. Error bars represent the mean ± SD of at least three independent experiments, p < .05, p < .01, p < .001.

Article Snippet: The membrane was then probed with primary antibodies at 4 C overnight and with species-specific secondary antibodies at room temperature for 1 h. The primary antibodies, including FOS antibody (1:1000, Proteintech, 66590-1-Ig), NF-jB antibody (1:1000, Proteintech, 14220-1-AP), MAPK8 antibody (1:1000, Proteintech, 66210-1-Ig), CREB5 antibody (1:1000, Proteintech, 14196-1-AP), GAPDH antibody (1:5000, Proteintech, 60004-1-Ig), CD9 antibody (1:1000, Abcam, ab223052), CD63 antibody (1:1000, Abcam, ab68418), TSG101 Figure 1.

Techniques: Expressing, Inhibition, Incubation, Produced, Concentration Assay, Derivative Assay, Transmission Assay, Microscopy

Journal: Cell Death & Disease

Article Title: Airway epithelial cGAS inhibits LPS-induced acute lung injury through CREB signaling

doi: 10.1038/s41419-023-06364-0

Figure Lengend Snippet: A The heat map of the differentially expressed genes (DEGs) (top 20 up-regulated and downregulate genes) in control- or cGAS-siRNA HBE cells through RNA sequencing analysis. Blue indicates a relatively low expression and red indicates a relatively high expression. B The volcano plot of differentially expressed genes (DEGs) in control- or cGAS-siRNA HBE cells through RNA sequencing analysis. C RT-qPCR of CREB in HBE cells transfected with cGAS siRNA or control siRNA for 24 h and incubated with 100 μg/ml LPS for an additional 24 h ( n = 3). Representative immunofluorescence images ( D ) and quantification of staining ( E ) of P-CREB (red) and nuclei (DAPI, blue) in HBE cells transfected with cGAS siRNA or control siRNA for 24 h and incubated with 100 μg/ml LPS for an additional 24 h. Scale bars, 10 μm. F HBE cells were transfected with cGAS siRNA or control siRNA for 24 h and were treated with LPS (100 μg/ml) for 24 h, and the cell lysates were collected for immunoblotting. G Representative lung immunofluorescence images of P-CREB (red), nuclei (DAPI, blue), and airway epithelial cells (marked with CC10, green) in mice. Scale bars, 100 μm. Two-way ANOVA with Sidaks multiple comparisons ( C , E ), *** p < 0.001, **** p < 0.0001. The data presented are one representative experiment of three independent experiments and shown as mean ± SEM.

Article Snippet: SiRNA of CREB (SR306377) was purchased from Origene (USA).

Techniques: Control, RNA Sequencing Assay, Expressing, Quantitative RT-PCR, Transfection, Incubation, Immunofluorescence, Staining, Western Blot

Journal: Cell Death & Disease

Article Title: Airway epithelial cGAS inhibits LPS-induced acute lung injury through CREB signaling

doi: 10.1038/s41419-023-06364-0

Figure Lengend Snippet: A – D HBE cells transfected with cGAS-, CREB- or control-siRNA were incubated with 100 μg/ml LPS for 24 h. RT-qPCR ( A , B ) and ELISA ( C , D ) analysis of IL-6 and IL-8 of HBE cells and culture supernatants were performed. The data presented are one representative experiment of three independent experiments and shown as mean ± SEM and statistical analyses were calculated using two-way ANOVA with Sidaks multiple comparisons. E , F CC10-rtTA/(tetO)7-Cre/cGAS flox/flox mice (KO) and their littermate controls (WT) were fed with doxycycline in their drinking water (2 mg/ml) for 20 days. The mice were pretreated with 10 mg/kg 666-15(CREB inhibitor) or DMSO for 2 h and then intratracheally challenged with LPS (5 mg/kg) for 24 h. E The count of the inflammatory cells in BALF from mice. F RT-qPCR of IL-6, IL-1β, TNF-α, CXCL1, and CXCL2 in lung tissues from mice. In E and F , each symbol represents an individual mouse ( n = 4~7). The data shown as mean ± SEM and statistical analyses were calculated using one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: SiRNA of CREB (SR306377) was purchased from Origene (USA).

Techniques: Transfection, Control, Incubation, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Epigenetically regulated miR-449a enhances hepatitis B virus replication by targeting cAMP-responsive element binding protein 5 and modulating hepatocytes phenotype

doi: 10.1038/srep25389

Figure Lengend Snippet: (A) HepG2.2.15 cells were transfected with 20 nM of different siRNAs targeting MMAB, GAL, DDI2, TMEM194B, CREB5, MCM10, SASS6, MYBL1, ZMAT1 , and APOBEC3B for 4 days. HBV replication was detected by Southern blotting. ( B ) Western blot analysis of CREB5 and FXRα protein expression in HepG2.2.15 cells transfected with miR-449a or siRNA specific for CREB5 at 20 nM for 3 days. ( C ) Wildtype and mutated pmiR-CREB5-3UTR luciferase reporters (100 ng each) were co-transfected with 20 nM of miR-449a in Huh7 cells, luciferase activity was assayed at 48 h. The fold change of luciferase expression expressed as the ratio of miR-449a- to miR-con-transfected samples. ( D ) The CREB5 expression vector pcDNA3.1/V5-CREB5 was co-transfected in Huh7 cells with pSM2 at the indicated concentrations for 4 days. HBV replication was detected by Southern blotting, and CREB5 expression was determined by V5-tag western blotting. The levels of secreted HBsAg and HBeAg in culture media were measured by the CMIA test. *P < 0.05.

Article Snippet: Protein samples were subjected to SDS-PAGE, blotted and then probed with primary antibodies against the following: ALB, CDK6, E2F1, HDAC1, ERK1/2, phosphorylated-Rb and -ERK1/2 (Cell Signaling Technology, Danvers, MA); CREB5 and FXRα (R&D System, Minneapolis, MN); and β-actin (Sigma-Aldrich).

Techniques: Transfection, Southern Blot, Western Blot, Expressing, Luciferase, Activity Assay, Plasmid Preparation

Journal: Molecules and Cells

Article Title: Neurotoxin-Induced Pathway Perturbation in Human Neuroblastoma SH-EP Cells

doi: 10.14348/molcells.2014.0173

Figure Lengend Snippet: Target genes for real-timeqPCR and their corresponding TaqMan Gene Expression Assay IDs

Article Snippet: CREB5 , Hs00329596_s1 , cAMP responsive element binding protein 5 (CREB5), transcript variant 1, mRNA.

Techniques: Gene Expression, Variant Assay, Binding Assay

Journal: Molecules and Cells

Article Title: Neurotoxin-Induced Pathway Perturbation in Human Neuroblastoma SH-EP Cells

doi: 10.14348/molcells.2014.0173

Figure Lengend Snippet: Average fold changes for 21 genes obtained from microarray and real-time qPCR at 3 and 24 h after MPP + treatment

Article Snippet: CREB5 , Hs00329596_s1 , cAMP responsive element binding protein 5 (CREB5), transcript variant 1, mRNA.

Techniques: Microarray

Journal: Oncology Letters

Article Title: CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer

doi: 10.3892/ol.2017.7234

Figure Lengend Snippet: cAMP response element-binding protein 5 (CREB5) is negatively associated with ovarian cancer prognosis. (A) CREB5 expression profiling from GSE41 2 datasets (n=46). (B) Overall survival of patients with low vs. high CREB5 expression in the GSE4122 dataset (n=580). (C) Relapse-free survival of patients with low vs. high CREB5 expression in the GSE4122 dataset (n=484).

Article Snippet: After blocking, the membranes were incubated with anti-CREB5 monoclonal antibodies (1:1,000; GTX44660; BD Biosciences, Franklin Lakes, NJ, USA) overnight at 4°C.

Techniques: Binding Assay, Expressing

Journal: Oncology Letters

Article Title: CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer

doi: 10.3892/ol.2017.7234

Figure Lengend Snippet: (A and B) qPCR analysis of CREB5 expression in (B) 10 ovarian cancer cell lines and (A) 10 ovarian cancer samples compared to human ovarian surface epithelial cells (HOSEpiC) and IOSE80. Transcript levels were normalized to GAPDH expression. Bars represent the mean ± standard deviation of three independent experiments. *P<0.05. (C) Western blot analysis of CREB5 protein expression in normal ovarian epithelial cells (HOSEpiC and IOSE80) and primary ovarian tumors from 10 patients with ovarian cancer (T1-T10). GAPDH was used as the loading control. (D) Western blot assay of CREB5 protein expression in normal ovarian epithelial cells and ovarian cancer cell lines. IOSE80 cells were used as the negative control.

Article Snippet: After blocking, the membranes were incubated with anti-CREB5 monoclonal antibodies (1:1,000; GTX44660; BD Biosciences, Franklin Lakes, NJ, USA) overnight at 4°C.

Techniques: Expressing, Standard Deviation, Western Blot, Negative Control

Journal: Oncology Letters

Article Title: CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer

doi: 10.3892/ol.2017.7234

Figure Lengend Snippet: (A) Immunohistochemical staining of CREB5 in human ovarian epithelial cancer tissues. *P<0.05. (B) Overall survival of patients with low vs. high expression of CREB5 in human ovarian surface epithelial cells (P<0.01). (C) Relapse-free survival (RFS) of patients with low vs. high CREB5 expression in ovarian cancer cells (P<0.01).

Article Snippet: After blocking, the membranes were incubated with anti-CREB5 monoclonal antibodies (1:1,000; GTX44660; BD Biosciences, Franklin Lakes, NJ, USA) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Oncology Letters

Article Title: CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer

doi: 10.3892/ol.2017.7234

Figure Lengend Snippet: The relationship between CREB5 and clinical pathological characteristics in 125 patients with ovarian cancer.

Article Snippet: After blocking, the membranes were incubated with anti-CREB5 monoclonal antibodies (1:1,000; GTX44660; BD Biosciences, Franklin Lakes, NJ, USA) overnight at 4°C.

Techniques: Expressing

Journal: Oncology Letters

Article Title: CREB5 promotes tumor cell invasion and correlates with poor prognosis in epithelial ovarian cancer

doi: 10.3892/ol.2017.7234

Figure Lengend Snippet: Univariate and multivariate analysis of factors associated with overall survival in 125 ovarian cancer patients.

Article Snippet: After blocking, the membranes were incubated with anti-CREB5 monoclonal antibodies (1:1,000; GTX44660; BD Biosciences, Franklin Lakes, NJ, USA) overnight at 4°C.

Techniques: