Journal: The FASEB Journal

Article Title: Prolonged activation of cAMP signaling leads to endothelial barrier disruption via transcriptional repression of RRAS

doi: 10.1096/fj.201700818RRR

Figure Lengend Snippet: RNA sequence analysis of human lung microvascular ECs

Article Snippet: Expression vectors pCMV-N-Flag and pCMV-N-Flag-CREB1 encoding a full-length cDNA for human CREB1 flanked with an N-terminal Flag tag were purchased from Sino Biological (Beijing, China).

Techniques: Sequencing

Journal: Journal of affective disorders

Article Title: Alteration of Cyclic-AMP Response Element Binding Protein in the Postmortem Brain of Subjects with Bipolar Disorder and Schizophrenia

doi: 10.1016/j.jad.2013.09.033

Figure Lengend Snippet: A. Representative Western blots showing the immunolabeling of CREB and β-actin in the membrane fraction of DLPFC and CG of two normal controls (NC), two schizophrenic (SZ), and two bipolar (BP) subjects. kDa indicates kilo Daltons.

Article Snippet: Briefly, 1 μg of total RNA was reverse transcribed using 50 ng random hexamers, 2mM dNTP mix, 10 units ribonuclease inhibitor, and 200 units MMLV-reverse transcriptase enzyme in a final reaction volume of 20 μl. qRT-PCR was performed with MX3005p sequence detection system (Agilent) using pre-designed Taqman gene expression assays (Applied Biosystems, Foster City, CA) targeting CREB1, Hs00231713_m1 two housekeeping genes β-actin (ACTB), Hs99999903_m1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and Hs99999905_m1.

Techniques: Western Blot, Immunolabeling, Membrane

Journal: Journal of affective disorders

Article Title: Alteration of Cyclic-AMP Response Element Binding Protein in the Postmortem Brain of Subjects with Bipolar Disorder and Schizophrenia

doi: 10.1016/j.jad.2013.09.033

Figure Lengend Snippet: The mean mRNA expression levels of CREB in the DLPFC of NC 1.00 ± 0.09 (n = 20), SZ 0.86 ± 0.10 (n = 20), and BP 0.71 ± 0.08 (n = 19) subjects.

Article Snippet: Briefly, 1 μg of total RNA was reverse transcribed using 50 ng random hexamers, 2mM dNTP mix, 10 units ribonuclease inhibitor, and 200 units MMLV-reverse transcriptase enzyme in a final reaction volume of 20 μl. qRT-PCR was performed with MX3005p sequence detection system (Agilent) using pre-designed Taqman gene expression assays (Applied Biosystems, Foster City, CA) targeting CREB1, Hs00231713_m1 two housekeeping genes β-actin (ACTB), Hs99999903_m1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and Hs99999905_m1.

Techniques: Expressing

Journal: British Journal of Nutrition

Article Title: Effect of vitamin D3 in reducing metabolic and oxidative stress in the liver of streptozotocin-induced diabetic rats

doi: 10.1017/s0007114511006830

Figure Lengend Snippet: Fig. 2. Real-time amplification of insulin receptor (INSR) ( ), GLUT2 ( ), phospholipase C (PLC, ), cyclic AMP-responsive element-binding protein (CREB, ) and vitamin D receptor (VDR, ) mRNA from the liver of the experimental rats. Values are means of six to eight separate experiments (n 8–10 animals per group), with standard errors of the mean represented by vertical bars. * Mean values were significantly different when compared with the control group (P,0·05). † Mean values were significantly different when compared with the diabetic group. RQ, relative quantification; D þ I, insulin-treated diabetic rats; D þ V, vitamin D3-treated diabetic rats.

Article Snippet: The specific primers of GPx (Rn00577994_g1), SOD (Rn01477289_m1), insulin receptor (INSR) (Rn00567070_m1), GLUT2 (Rn00563565_m1), PLC (Rn01647142_m1), CREB (Rn00578826_m1) and VDR (Rn00566976_m1) were purchased from Applied Biosystems.

Techniques: Binding Assay, Control

Journal: bioRxiv

Article Title: Dibutyryl cyclic AMP downregulates tenascin-C in neurons and astrocytes and reduces AAV-mediated gene expression in DRG neurons

doi: 10.1101/2025.05.13.653846

Figure Lengend Snippet: ( A ) Representative snapshots from Imaris software of DRGs visualized using the Zeiss Lightsheet Z.1 following ECi clearing. Scale bar: 400 µm. ( B ) Representative confocal images of DRG sections showing varying signal intensities in the presence and absence of db-cAMP. For intensity measurements, all stained sections were imaged using the same laser power, gain, and airy units. Scale bar: 400 µm. ( C ) Quantification of (A). Bar graphs display the total number of GFP-positive or V5-positive cells in the DRG. Data are presented as mean ± SEM, analysed by unpaired t-test, ns > 0.5, *p < 0.05, n = 6 DRGs/group. ( D ) Quantification of (B). Bar graphs show the total GFP-positive or V5-positive signal per DRG. Data are presented as mean ± SEM, analysed by unpaired t-test, **p < 0.01, ****p < 0.0001, n = 8 DRGs/group, with three sections evaluated per DRG to obtain the final value. ( E, F ) Immunoblotting results for GFP, PKA C-α, β-actin, and vinculin in DRG samples from animals injected with GFP only and those co-injected with db-cAMP 13 weeks post-injection. Values were normalised to vinculin. Two housekeeping proteins were utilised to enhance the reliability and accuracy of the results. Results are presented as mean density values ± SEM; n = 3 DRGs, with western blots repeated four times in separate experiments; ns > 0.5, *p < 0.05, **p < 0.01, analysed by unpaired t-test. ( G ) Heat maps illustrate Log 2 fold changes (as Log 2 (2^-ΔΔCt)) for the expression of Creb1 and Itg α 9 , normalised to the uninjured, uninjected control. Log 2 (2^-ΔΔCt) values were determined through qRT-PCR analysis. Results are presented as a heat map.

Article Snippet: For qPCR, TaqMan® gene expression assays (Life Technologies by Thermo Fisher Scientific, Waltham, MA, USA) were used for CREB1 (Rn00578828_g1), Itga9 (Rn01746751_m1) and GAPDH (Rn01775763_g1), all purchased from Applied Biosystems and used according to the manufacturer’s recommendations.

Techniques: Software, Staining, Western Blot, Injection, Expressing, Control, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Macrophage migration inhibitory factor mediates joint capsule fibrosis via facilitating phospholipid metabolite PGE2 production in fibroblasts

doi: 10.1007/s00018-025-05800-y

Figure Lengend Snippet: Effects of CREB on the MIF-induced COX2 expression and PGE2 production. A , Western blot analysis of p-CREB following joint capsule fibroblasts treated with 2 µg/mL MIF for 0, 15, 30, 60, and 120 min. B , Quantitative results of A. C , Western blot analysis of p-CREB in joint capsule fibroblasts following siRNA2 knockdown of CD74 for 48 h and 2 µg/mL MIF stimulation for 15 min. D , Quantitative results of C. E , Evaluation of CREB knockdown efficiency using qRT-PCR after transfection for 48 h and siRNA1 was selected for subsequent experiments. F–H , Following siRNA1 knockdown of CREB for 48 h and 2 μg/ml MIF stimulation for 24 h, COX2 expression in joint capsule fibroblasts was assessed by qRT-PCR ( F ) and Western blot ( G ). Quantitative results of G as shown in ( H ). I and J , PGE2 production in supernatant ( I ) and lysate ( J ) of joint capsule fibroblasts were tested by ELISA accordingly. Each experiment was conducted three times. Error bars denote standard deviation. * P < 0.05 compared with Control group or 0 min group

Article Snippet: CREB gene silencing (RiboBio, Guangzhou, China) was similarly performed using different siRNA sequences (siRNA1, siRNA2, siRNA3) following the same procedures.

Techniques: Expressing, Western Blot, Knockdown, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control