Review

creb inhibitor compound (Tocris)

Tocris is a verified supplier

Tocris manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Tocris creb inhibitor compound

(A) Schematic representation of the <t>CREB</t> reporter used in the CRISPR-based screen. Two cAMP response elements were fused to a ProteoTuner destabilizing domain (degron) followed by a green fluorescent protein, and inserted into a lentiviral vector. A clonal cell line was generated by transducing HEK293 cells with the reporter and dCas9-BFP-KRAB, sorting individual cells, and growing and verifying clonal lines for high reporter expression and efficient dCas9-dependent gene silencing. (B) The CREB reporter responds robustly to direct stimulation of adenylyl cyclase/cAMP signaling with forskolin. Reporter cells were treated with 10 μM forskolin (FSK) <t>or</t> <t>DMSO</t> (vehicle), and 1 μM Shield-1 was added simultaneously to stabilize the degron domain. After 4 h, reporter expression was analyzed by flow cytometry. Data from n = 4 per condition. (C) The CREB and PKA inhibitors, 666–15 and H89, respectively, significantly diminish FSK-induced accumulation of the reporter. Cells were pre-incubated with 100 nM 666–15 or 10 μM H89 versus DMSO (vehicle) for 30 min, then treated with 10 μM FSK in the presence of 1 μM Shield-1. After 4 h, reporter accumulation was analyzed by flow cytometry. Data plotted are from n = 5 for 666–15 and n = 7 for H89. (D) Timecourse for β2-AR-dependent reporter induction. Reporter cells were treated with 1 μM isoproterenol / 1 μM Shield-1 for indicated times or treated with 1 μM Shield-1 alone (no isoproterenol) for 5 h (“untreated”), and reporter expression was analyzed by flow cytometry. Data from n = 5. (E) Dose-response curve for isoproterenol-dependent reporter expression. Reporter cells were treated with indicated doses of isoproterenol and 1 μM Shield-1 for 4 h, and reporter expression was analyzed by flow cytometry. Data plotted are means of GFP expression, n = 3 per condition. EC 50 curve-fitting was performed using Prism6 GraphPad software. In each flow cytometry experiment, 10,000 cells total were analyzed and gated for singlets. Error bars = ± s.e.m. **** = p ≤ 0.0001; ** = p ≤ 0.01 by unpaired two-tailed Student’s t-test.

Creb Inhibitor Compound, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/creb inhibitor compound/product/Tocris
Average 93 stars, based on 40 article reviews

creb inhibitor compound - by Bioz Stars, 2026-03

93/100 stars

Images

1) Product Images from "A high-throughput CRISPR interference screen for dissecting functional regulators of GPCR/cAMP signaling"

Article Title: A high-throughput CRISPR interference screen for dissecting functional regulators of GPCR/cAMP signaling

Journal: PLoS Genetics

doi: 10.1371/journal.pgen.1009103

(A) Schematic representation of the CREB reporter used in the CRISPR-based screen. Two cAMP response elements were fused to a ProteoTuner destabilizing domain (degron) followed by a green fluorescent protein, and inserted into a lentiviral vector. A clonal cell line was generated by transducing HEK293 cells with the reporter and dCas9-BFP-KRAB, sorting individual cells, and growing and verifying clonal lines for high reporter expression and efficient dCas9-dependent gene silencing. (B) The CREB reporter responds robustly to direct stimulation of adenylyl cyclase/cAMP signaling with forskolin. Reporter cells were treated with 10 μM forskolin (FSK) or DMSO (vehicle), and 1 μM Shield-1 was added simultaneously to stabilize the degron domain. After 4 h, reporter expression was analyzed by flow cytometry. Data from n = 4 per condition. (C) The CREB and PKA inhibitors, 666–15 and H89, respectively, significantly diminish FSK-induced accumulation of the reporter. Cells were pre-incubated with 100 nM 666–15 or 10 μM H89 versus DMSO (vehicle) for 30 min, then treated with 10 μM FSK in the presence of 1 μM Shield-1. After 4 h, reporter accumulation was analyzed by flow cytometry. Data plotted are from n = 5 for 666–15 and n = 7 for H89. (D) Timecourse for β2-AR-dependent reporter induction. Reporter cells were treated with 1 μM isoproterenol / 1 μM Shield-1 for indicated times or treated with 1 μM Shield-1 alone (no isoproterenol) for 5 h (“untreated”), and reporter expression was analyzed by flow cytometry. Data from n = 5. (E) Dose-response curve for isoproterenol-dependent reporter expression. Reporter cells were treated with indicated doses of isoproterenol and 1 μM Shield-1 for 4 h, and reporter expression was analyzed by flow cytometry. Data plotted are means of GFP expression, n = 3 per condition. EC 50 curve-fitting was performed using Prism6 GraphPad software. In each flow cytometry experiment, 10,000 cells total were analyzed and gated for singlets. Error bars = ± s.e.m. **** = p ≤ 0.0001; ** = p ≤ 0.01 by unpaired two-tailed Student’s t-test.

Figure Legend Snippet: (A) Schematic representation of the CREB reporter used in the CRISPR-based screen. Two cAMP response elements were fused to a ProteoTuner destabilizing domain (degron) followed by a green fluorescent protein, and inserted into a lentiviral vector. A clonal cell line was generated by transducing HEK293 cells with the reporter and dCas9-BFP-KRAB, sorting individual cells, and growing and verifying clonal lines for high reporter expression and efficient dCas9-dependent gene silencing. (B) The CREB reporter responds robustly to direct stimulation of adenylyl cyclase/cAMP signaling with forskolin. Reporter cells were treated with 10 μM forskolin (FSK) or DMSO (vehicle), and 1 μM Shield-1 was added simultaneously to stabilize the degron domain. After 4 h, reporter expression was analyzed by flow cytometry. Data from n = 4 per condition. (C) The CREB and PKA inhibitors, 666–15 and H89, respectively, significantly diminish FSK-induced accumulation of the reporter. Cells were pre-incubated with 100 nM 666–15 or 10 μM H89 versus DMSO (vehicle) for 30 min, then treated with 10 μM FSK in the presence of 1 μM Shield-1. After 4 h, reporter accumulation was analyzed by flow cytometry. Data plotted are from n = 5 for 666–15 and n = 7 for H89. (D) Timecourse for β2-AR-dependent reporter induction. Reporter cells were treated with 1 μM isoproterenol / 1 μM Shield-1 for indicated times or treated with 1 μM Shield-1 alone (no isoproterenol) for 5 h (“untreated”), and reporter expression was analyzed by flow cytometry. Data from n = 5. (E) Dose-response curve for isoproterenol-dependent reporter expression. Reporter cells were treated with indicated doses of isoproterenol and 1 μM Shield-1 for 4 h, and reporter expression was analyzed by flow cytometry. Data plotted are means of GFP expression, n = 3 per condition. EC 50 curve-fitting was performed using Prism6 GraphPad software. In each flow cytometry experiment, 10,000 cells total were analyzed and gated for singlets. Error bars = ± s.e.m. **** = p ≤ 0.0001; ** = p ≤ 0.01 by unpaired two-tailed Student’s t-test.

Techniques Used: CRISPR, Plasmid Preparation, Generated, Expressing, Flow Cytometry, Incubation, Software, Two Tailed Test

Image Search Results

Journal: PLoS Genetics

Article Title: A high-throughput CRISPR interference screen for dissecting functional regulators of GPCR/cAMP signaling

doi: 10.1371/journal.pgen.1009103

Figure Lengend Snippet: (A) Schematic representation of the CREB reporter used in the CRISPR-based screen. Two cAMP response elements were fused to a ProteoTuner destabilizing domain (degron) followed by a green fluorescent protein, and inserted into a lentiviral vector. A clonal cell line was generated by transducing HEK293 cells with the reporter and dCas9-BFP-KRAB, sorting individual cells, and growing and verifying clonal lines for high reporter expression and efficient dCas9-dependent gene silencing. (B) The CREB reporter responds robustly to direct stimulation of adenylyl cyclase/cAMP signaling with forskolin. Reporter cells were treated with 10 μM forskolin (FSK) or DMSO (vehicle), and 1 μM Shield-1 was added simultaneously to stabilize the degron domain. After 4 h, reporter expression was analyzed by flow cytometry. Data from n = 4 per condition. (C) The CREB and PKA inhibitors, 666–15 and H89, respectively, significantly diminish FSK-induced accumulation of the reporter. Cells were pre-incubated with 100 nM 666–15 or 10 μM H89 versus DMSO (vehicle) for 30 min, then treated with 10 μM FSK in the presence of 1 μM Shield-1. After 4 h, reporter accumulation was analyzed by flow cytometry. Data plotted are from n = 5 for 666–15 and n = 7 for H89. (D) Timecourse for β2-AR-dependent reporter induction. Reporter cells were treated with 1 μM isoproterenol / 1 μM Shield-1 for indicated times or treated with 1 μM Shield-1 alone (no isoproterenol) for 5 h (“untreated”), and reporter expression was analyzed by flow cytometry. Data from n = 5. (E) Dose-response curve for isoproterenol-dependent reporter expression. Reporter cells were treated with indicated doses of isoproterenol and 1 μM Shield-1 for 4 h, and reporter expression was analyzed by flow cytometry. Data plotted are means of GFP expression, n = 3 per condition. EC 50 curve-fitting was performed using Prism6 GraphPad software. In each flow cytometry experiment, 10,000 cells total were analyzed and gated for singlets. Error bars = ± s.e.m. **** = p ≤ 0.0001; ** = p ≤ 0.01 by unpaired two-tailed Student’s t-test.

Article Snippet: The CREB inhibitor compound, 666–15, was purchased from Tocris Bioscience (Cat #5661), resuspended in DMSO and used at 100 nM final.

Techniques: CRISPR, Plasmid Preparation, Generated, Expressing, Flow Cytometry, Incubation, Software, Two Tailed Test

Knockdown of PAK4 triggers neurodegeneration and inhibits CREB pathway. A, Western blotting analysis of the efficiency of PAK4 knockdown using siRNAs. The mRNA level of PAK4 was detected by qRT‐PCR in NSC34 cells transfected with siNC or siPAK4. B, Flow cytometry detected the apoptosis of NSC34 cells after transfection with siPAK4‐2 or siNC for 72 h. C, Quantification of cell apoptosis. D and E, TUNEL staining of NSC34 cells after treatment with siPAK4‐2. F, Western blotting of PAK4/CREB pathway signalling proteins expression levels in PAK4 knockdown NSC34 cells. G, Quantification of the blots in (F) normalized to β‐actin. Data were presented as means ± SD. * P < .05, ** P < .01, *** P < .001. Student's t ‐test (C, E, G) or ANOVA with Student‐Newman‐Keuls post hoc analysis (A). Scale bar = 50 μm

Journal: Cell Proliferation

Article Title: PAK4 suppresses motor neuron degeneration in hSOD1 G93A ‐linked amyotrophic lateral sclerosis cell and rat models

doi: 10.1111/cpr.13003

Figure Lengend Snippet: Knockdown of PAK4 triggers neurodegeneration and inhibits CREB pathway. A, Western blotting analysis of the efficiency of PAK4 knockdown using siRNAs. The mRNA level of PAK4 was detected by qRT‐PCR in NSC34 cells transfected with siNC or siPAK4. B, Flow cytometry detected the apoptosis of NSC34 cells after transfection with siPAK4‐2 or siNC for 72 h. C, Quantification of cell apoptosis. D and E, TUNEL staining of NSC34 cells after treatment with siPAK4‐2. F, Western blotting of PAK4/CREB pathway signalling proteins expression levels in PAK4 knockdown NSC34 cells. G, Quantification of the blots in (F) normalized to β‐actin. Data were presented as means ± SD. * P < .05, ** P < .01, *** P < .001. Student's t ‐test (C, E, G) or ANOVA with Student‐Newman‐Keuls post hoc analysis (A). Scale bar = 50 μm

Article Snippet: At 48 hours after transfection, cells containing cDNA or vehicle control were incubated with 50.26 ng/mL CREB inhibitor (compound 3i, Selleckchem, US).

Techniques: Knockdown, Western Blot, Quantitative RT-PCR, Transfection, Flow Cytometry, TUNEL Assay, Staining, Expressing

CREB pathway mediates PAK4‐induced neuroprotection. A, Immunoblotting of cell extract from EV, EV + 3i, PAK4 + and PAK4 + +3i groups. B‐F, Quantitative analysis of PAK4 (B), CREB (C), pCREB (D), PGC‐1a (E) and Bcl‐2 (F) levels in (A) normalized to β‐actin. Data were presented as means ± SD. ns ≥ .05, * P < .05, ** P < .01, *** P < .001, vs Control group. ANOVA with Student‐Newman‐Keuls post hoc analysis