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atypical pkcs inhibitor crt0066854 hydrochloride (Tocris)

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Tocris atypical pkcs inhibitor crt0066854 hydrochloride

Figure 11. Involvement of PKCζ in the cytostatic and morphological activities of huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated for 30 min with 2.5 µM PKCζ inhibitor <t>(CRT0066854</t> hydrochloride) and then incubated with daphnanes at 1 µg/mL. A) After 48 h of incubation, cell viability was measured by colorimetry and the activity of daphnanes was calculated relative to the DMSO control (Caco-2 cells treated with DMSO solvent only). Mean ± S.E.M. from 3 (n) independent experiments and t-test compared to DMSO controls in the presence or absence of PKCζ inhibitor: *p < 0.05. B) Morphological impact of Caco-2 cells pre- treatment with the PKCζ inhibitor before addition of huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT). Nuclei (cyan) and cytoskeleton (red) labeling (confocal views taken at the OPERA Phenix®, 20X objective, size scale = 50 µm). Photographs are representative of duplicates from one experiment. C) After 48 h of incubation cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pThr410) and the total form of PKCζ are shown. Actin shows the equivalence of protein deposition. The modulation of the phosphorylated active form of PKCζ (pThr410) after addition of the PKCζ inhibitor is shown. Quantification of n = 2 experiments and statistical t-test compared to DMSO are shown: Mean ± S.E.M., *p < 0.05.

Atypical Pkcs Inhibitor Crt0066854 Hydrochloride, supplied by Tocris, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 91 stars, based on 3 article reviews

atypical pkcs inhibitor crt0066854 hydrochloride - by Bioz Stars, 2026-10

91/100 stars

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1) Product Images from "Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells."

Article Title: Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells.

Journal: Bioorganic & medicinal chemistry

doi: 10.1016/j.bmc.2023.117366

Figure Legend Snippet: Figure 11. Involvement of PKCζ in the cytostatic and morphological activities of huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated for 30 min with 2.5 µM PKCζ inhibitor (CRT0066854 hydrochloride) and then incubated with daphnanes at 1 µg/mL. A) After 48 h of incubation, cell viability was measured by colorimetry and the activity of daphnanes was calculated relative to the DMSO control (Caco-2 cells treated with DMSO solvent only). Mean ± S.E.M. from 3 (n) independent experiments and t-test compared to DMSO controls in the presence or absence of PKCζ inhibitor: *p < 0.05. B) Morphological impact of Caco-2 cells pre- treatment with the PKCζ inhibitor before addition of huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT). Nuclei (cyan) and cytoskeleton (red) labeling (confocal views taken at the OPERA Phenix®, 20X objective, size scale = 50 µm). Photographs are representative of duplicates from one experiment. C) After 48 h of incubation cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pThr410) and the total form of PKCζ are shown. Actin shows the equivalence of protein deposition. The modulation of the phosphorylated active form of PKCζ (pThr410) after addition of the PKCζ inhibitor is shown. Quantification of n = 2 experiments and statistical t-test compared to DMSO are shown: Mean ± S.E.M., *p < 0.05.

Techniques Used: Incubation, Colorimetric Assay, Activity Assay, Control, Solvent, Labeling, Western Blot

Figure 13. Regulation of GSK3β, Akt and β-catenin by huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated or not for 30 min with 2.5 µM PKCζ in hibitor (CRT0066854 hydrochloride) prior to huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT) treatment (1 µg/mL). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated forms (pSer9-GSK3β; p-Ser473-Akt) and the total forms of GSK3β, Akt and β-catenin are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO (Ctrl) are shown: Mean ± S.E.M., *p < 0.05, **p < 0.01.

Figure Legend Snippet: Figure 13. Regulation of GSK3β, Akt and β-catenin by huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated or not for 30 min with 2.5 µM PKCζ in hibitor (CRT0066854 hydrochloride) prior to huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT) treatment (1 µg/mL). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated forms (pSer9-GSK3β; p-Ser473-Akt) and the total forms of GSK3β, Akt and β-catenin are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO (Ctrl) are shown: Mean ± S.E.M., *p < 0.05, **p < 0.01.

Techniques Used: Incubation, Western Blot

Figure 14. Implication of MEK in the cytostatic effects of mono-esterified daphnanes. A) Huratoxin (1 µg/mL) was incubated with Caco-2 cells for 48 h before cell viability test. Prior addition of huratoxin, a MEK inhibitor (PD0325901) was added or not for 15 min at the concentration of 0.5 µM. Data are from n = 3 experiments, Mean ± S.E.M., statistical t-test compared to DMSO (Ctrl) are shown: ***p < 0.001. B) Huratoxin (HT) and 4′,5′-epoxy-huratoxin (EHT) at 1 µg/mL were incubated with Caco-2 cells which have been pretreated or not for 30 min with 2.5 µM PKCζ inhibitor (CRT0066854 hydrochloride). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pSer217/221-MEK) and the total form of MEK are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO(Ctrl) are shown: Mean ± S.E.M., *p < 0.05.

Figure Legend Snippet: Figure 14. Implication of MEK in the cytostatic effects of mono-esterified daphnanes. A) Huratoxin (1 µg/mL) was incubated with Caco-2 cells for 48 h before cell viability test. Prior addition of huratoxin, a MEK inhibitor (PD0325901) was added or not for 15 min at the concentration of 0.5 µM. Data are from n = 3 experiments, Mean ± S.E.M., statistical t-test compared to DMSO (Ctrl) are shown: ***p < 0.001. B) Huratoxin (HT) and 4′,5′-epoxy-huratoxin (EHT) at 1 µg/mL were incubated with Caco-2 cells which have been pretreated or not for 30 min with 2.5 µM PKCζ inhibitor (CRT0066854 hydrochloride). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pSer217/221-MEK) and the total form of MEK are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO(Ctrl) are shown: Mean ± S.E.M., *p < 0.05.

Techniques Used: Incubation, Concentration Assay, Western Blot

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Article Title: Discovery of lead natural products for developing pan-SARS-CoV-2 therapeutics.
Article Snippet: Bryostatin (2383), Gö6976 (2253), PEP005 (4054) and CRT 0066854 (5922) were obtained from Tocris Bioscience.

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Journal: Bioorganic & medicinal chemistry

Article Title: Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells.

doi: 10.1016/j.bmc.2023.117366

Figure Lengend Snippet: Figure 11. Involvement of PKCζ in the cytostatic and morphological activities of huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated for 30 min with 2.5 µM PKCζ inhibitor (CRT0066854 hydrochloride) and then incubated with daphnanes at 1 µg/mL. A) After 48 h of incubation, cell viability was measured by colorimetry and the activity of daphnanes was calculated relative to the DMSO control (Caco-2 cells treated with DMSO solvent only). Mean ± S.E.M. from 3 (n) independent experiments and t-test compared to DMSO controls in the presence or absence of PKCζ inhibitor: *p < 0.05. B) Morphological impact of Caco-2 cells pre- treatment with the PKCζ inhibitor before addition of huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT). Nuclei (cyan) and cytoskeleton (red) labeling (confocal views taken at the OPERA Phenix®, 20X objective, size scale = 50 µm). Photographs are representative of duplicates from one experiment. C) After 48 h of incubation cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pThr410) and the total form of PKCζ are shown. Actin shows the equivalence of protein deposition. The modulation of the phosphorylated active form of PKCζ (pThr410) after addition of the PKCζ inhibitor is shown. Quantification of n = 2 experiments and statistical t-test compared to DMSO are shown: Mean ± S.E.M., *p < 0.05.

Article Snippet: For some tests, atypical PKCs inhibitor CRT0066854 hydrochloride (0.625, 1.25 or 2.5 μM, Tocris Bioscience), PKCα inhibitor Gö6976 (0.2 μM, Calbiochem) and MEK inhibitor PD-0325901 (0.5 μM, MedChemExpress) were added 30 min (atypical PKCs inhibitor) or 15 min (PKCα and MEK inhibitors) before daphnanes (maximum concentration of DMSO 0,015%).

Techniques: Incubation, Colorimetric Assay, Activity Assay, Control, Solvent, Labeling, Western Blot

Journal: Bioorganic & medicinal chemistry

Article Title: Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells.

doi: 10.1016/j.bmc.2023.117366

Figure Lengend Snippet: Figure 13. Regulation of GSK3β, Akt and β-catenin by huratoxin and 4′,5′-epoxy-huratoxin. Caco-2 cells were pretreated or not for 30 min with 2.5 µM PKCζ in hibitor (CRT0066854 hydrochloride) prior to huratoxin (HT) or 4′,5′-epoxy-huratoxin (EHT) treatment (1 µg/mL). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated forms (pSer9-GSK3β; p-Ser473-Akt) and the total forms of GSK3β, Akt and β-catenin are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO (Ctrl) are shown: Mean ± S.E.M., *p < 0.05, **p < 0.01.

Techniques: Incubation, Western Blot

Journal: Bioorganic & medicinal chemistry

Article Title: Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells.

doi: 10.1016/j.bmc.2023.117366

Figure Lengend Snippet: Figure 14. Implication of MEK in the cytostatic effects of mono-esterified daphnanes. A) Huratoxin (1 µg/mL) was incubated with Caco-2 cells for 48 h before cell viability test. Prior addition of huratoxin, a MEK inhibitor (PD0325901) was added or not for 15 min at the concentration of 0.5 µM. Data are from n = 3 experiments, Mean ± S.E.M., statistical t-test compared to DMSO (Ctrl) are shown: ***p < 0.001. B) Huratoxin (HT) and 4′,5′-epoxy-huratoxin (EHT) at 1 µg/mL were incubated with Caco-2 cells which have been pretreated or not for 30 min with 2.5 µM PKCζ inhibitor (CRT0066854 hydrochloride). After 48 h of incubation, cells were lysed for Western blot analysis. The results obtained from the revelation of the phosphorylated form (pSer217/221-MEK) and the total form of MEK are shown. Actin shows the equivalence of protein deposition. Quantification of n = 2 experiments and statistical t-test compared to DMSO(Ctrl) are shown: Mean ± S.E.M., *p < 0.05.

Techniques: Incubation, Concentration Assay, Western Blot

(A and B) Human (A) and murine (B) GBM cell lines were assessed for expression of PKCι by immunoblot analysis using β-Actin as a loading control (upper panels) and for the effect of auranofin (middle panels) and CRT006854 (lower panels) on cell viability. Each point represents the mean ± 1 SD from 8 replicates. (C and D) Plot of auranofin (C) and CRT0066854 (D) EC 50 s versus PKCι expression in murine (red) and human (blue) GBM cell lines, fit to hyperbolic functions (solid red and blue lines). The significance (p value) is indicated in (C) and (D). Each point represents the mean ± 1 SD from 2–4 replicates. (E) IHC detection of PKCι in representative human PDX-derived GBM cell lines demonstrating low ( GBM8 and GBM43 , top) or high ( GBM12 and GBM10 , bottom) PKCι immunoscores. (F) Auranofin dose-response curves for low PKCι ( GBM8 and GBM43 , green curves) and high PKCι ( GBM12 and GBM10 , red curves) cell lines. Each point represents the mean ± 1 SD from 8 replicates. See also and .

Journal: Cell reports

Article Title: Protein kinase C ι and SRC signaling define reciprocally related subgroups of glioblastoma with distinct therapeutic vulnerabilities

doi: 10.1016/j.celrep.2021.110054

Figure Lengend Snippet: (A and B) Human (A) and murine (B) GBM cell lines were assessed for expression of PKCι by immunoblot analysis using β-Actin as a loading control (upper panels) and for the effect of auranofin (middle panels) and CRT006854 (lower panels) on cell viability. Each point represents the mean ± 1 SD from 8 replicates. (C and D) Plot of auranofin (C) and CRT0066854 (D) EC 50 s versus PKCι expression in murine (red) and human (blue) GBM cell lines, fit to hyperbolic functions (solid red and blue lines). The significance (p value) is indicated in (C) and (D). Each point represents the mean ± 1 SD from 2–4 replicates. (E) IHC detection of PKCι in representative human PDX-derived GBM cell lines demonstrating low ( GBM8 and GBM43 , top) or high ( GBM12 and GBM10 , bottom) PKCι immunoscores. (F) Auranofin dose-response curves for low PKCι ( GBM8 and GBM43 , green curves) and high PKCι ( GBM12 and GBM10 , red curves) cell lines. Each point represents the mean ± 1 SD from 8 replicates. See also and .

Article Snippet: CRT0066854 , Tocris Bioscience , Cat# 5922.

Techniques: Expressing, Western Blot, Control, Derivative Assay

(A) Trp53 −/− and Trp53 −/− / Prkci −/− GBM cells grown were treated with auranofin for 72 h. Each point represents the mean ± 1 SD from 10 replicates. (B and C) Trp53 −/− (B) and Trp53 −/− / Prkci −/− (C) cells were treated with auranofin ± 2 mM N -acetyl cysteine ( NAC ), and cell viability was assessed. Each point represents the mean ± 1 SD from 10 replicates. Data in (A)–(C) were fit to the Hill equation, constrained to eliminate data from the top and bottom 5% of drug concentrations to improve curve fitting. (D) Trp53 −/− and Trp53 −/− / Prkci −/− cells grown in 96-well plates were treated with CRT0066854 for 72 h. For (A)–(D), each point represents the mean ± 1 SD from 8 replicates. (E) Trp53 −/− cells (line 1) were treated with vehicle for 6 h or with 100 nM auranofin for 1, 3, and 6 h, and levels of total and phospho-ECT2 and ERK were measured by immunoblot (left panel). Band intensity is plotted as phospho-ERK/total ERK and phospho-ECT2/total ECT2 (right panel). Each point represents the mean ± 1 SD from 3 replicates. Significance was assessed by pairwise t test compared with the t = 0 vehicle control. (F) Invasion of Trp53 −/− cells through 3-μm Transwell membranes was measured in the presence of vehicle (closed) or 100 nM auranofin (open). Bar represents the mean ± 1 SD from 8 replicates. Significance was determined by two-tailed t test. See also and .

Journal: Cell reports

Article Title: Protein kinase C ι and SRC signaling define reciprocally related subgroups of glioblastoma with distinct therapeutic vulnerabilities

doi: 10.1016/j.celrep.2021.110054

Figure Lengend Snippet: (A) Trp53 −/− and Trp53 −/− / Prkci −/− GBM cells grown were treated with auranofin for 72 h. Each point represents the mean ± 1 SD from 10 replicates. (B and C) Trp53 −/− (B) and Trp53 −/− / Prkci −/− (C) cells were treated with auranofin ± 2 mM N -acetyl cysteine ( NAC ), and cell viability was assessed. Each point represents the mean ± 1 SD from 10 replicates. Data in (A)–(C) were fit to the Hill equation, constrained to eliminate data from the top and bottom 5% of drug concentrations to improve curve fitting. (D) Trp53 −/− and Trp53 −/− / Prkci −/− cells grown in 96-well plates were treated with CRT0066854 for 72 h. For (A)–(D), each point represents the mean ± 1 SD from 8 replicates. (E) Trp53 −/− cells (line 1) were treated with vehicle for 6 h or with 100 nM auranofin for 1, 3, and 6 h, and levels of total and phospho-ECT2 and ERK were measured by immunoblot (left panel). Band intensity is plotted as phospho-ERK/total ERK and phospho-ECT2/total ECT2 (right panel). Each point represents the mean ± 1 SD from 3 replicates. Significance was assessed by pairwise t test compared with the t = 0 vehicle control. (F) Invasion of Trp53 −/− cells through 3-μm Transwell membranes was measured in the presence of vehicle (closed) or 100 nM auranofin (open). Bar represents the mean ± 1 SD from 8 replicates. Significance was determined by two-tailed t test. See also and .

Article Snippet: CRT0066854 , Tocris Bioscience , Cat# 5922.

Techniques: Western Blot, Control, Two Tailed Test

(A and B) Trp53 −/− (blue) and Trp53 −/− /Prkci −/− (red) murine GBM cells were treated with saracatinib (A) and dasatinib (B) for 72 h. Each point represents the mean ± 1 SD from 8 replicates. (C) A plot of saracatinib EC 50 versus auranofin (ANF) EC 50 for eight human (red) and five murine (blue) GBM cell lines fit to rectangular hyperbolae (red and blue solid curves). Error bars indicate mean ± SEM. Statistical significance (p value) is indicated. (D and E) Dose-response curves of ANF (D) and saracatinib (E) in ANF-naive (closed symbols) and ANF-resistant (open symbols) Trp5 3 −/− GBM cells. Each point represents the mean ± 1 SD from 8 replicates. (F) (Top panel) Immunoblot of ANF-naive and -resistant cells for phosphorylated pY418 SRC and total SRC. (Bottom panel) Quantification of phospho-SRC for both ANF-naive and -resistant GBM cell lines. Results are expressed as pSRC/total SRC, and each point represents the mean ± 1 SD from 3 replicates. (G and H) Dose-response curves to ANF (G) and CRT0066854 (H) treatment of drug-naive and dasatinib-resistant MES1861 and MES4622 GBM cells. Each point represents the mean ± 1 SD from 8 replicates. (I) (Top panel) Representative immunoblots of lysates derived from three cultures of dasatinib-naive and -resistant MES1861cells for PKCι and actin. (Bottom panel) Quantification of PKCι expression by immunoblot analysis in dasatinib-naive and -resistant MES1861 and MES4622 GBM cells. Results are expressed as PKCι/β-Actin, and each point represents the mean ± 1 SD from 6 replicates. See also and .

Journal: Cell reports

Article Title: Protein kinase C ι and SRC signaling define reciprocally related subgroups of glioblastoma with distinct therapeutic vulnerabilities

doi: 10.1016/j.celrep.2021.110054

Figure Lengend Snippet: (A and B) Trp53 −/− (blue) and Trp53 −/− /Prkci −/− (red) murine GBM cells were treated with saracatinib (A) and dasatinib (B) for 72 h. Each point represents the mean ± 1 SD from 8 replicates. (C) A plot of saracatinib EC 50 versus auranofin (ANF) EC 50 for eight human (red) and five murine (blue) GBM cell lines fit to rectangular hyperbolae (red and blue solid curves). Error bars indicate mean ± SEM. Statistical significance (p value) is indicated. (D and E) Dose-response curves of ANF (D) and saracatinib (E) in ANF-naive (closed symbols) and ANF-resistant (open symbols) Trp5 3 −/− GBM cells. Each point represents the mean ± 1 SD from 8 replicates. (F) (Top panel) Immunoblot of ANF-naive and -resistant cells for phosphorylated pY418 SRC and total SRC. (Bottom panel) Quantification of phospho-SRC for both ANF-naive and -resistant GBM cell lines. Results are expressed as pSRC/total SRC, and each point represents the mean ± 1 SD from 3 replicates. (G and H) Dose-response curves to ANF (G) and CRT0066854 (H) treatment of drug-naive and dasatinib-resistant MES1861 and MES4622 GBM cells. Each point represents the mean ± 1 SD from 8 replicates. (I) (Top panel) Representative immunoblots of lysates derived from three cultures of dasatinib-naive and -resistant MES1861cells for PKCι and actin. (Bottom panel) Quantification of PKCι expression by immunoblot analysis in dasatinib-naive and -resistant MES1861 and MES4622 GBM cells. Results are expressed as PKCι/β-Actin, and each point represents the mean ± 1 SD from 6 replicates. See also and .

Article Snippet: CRT0066854 , Tocris Bioscience , Cat# 5922.

Techniques: Western Blot, Derivative Assay, Expressing

Journal: Cell reports

Article Title: Protein kinase C ι and SRC signaling define reciprocally related subgroups of glioblastoma with distinct therapeutic vulnerabilities

doi: 10.1016/j.celrep.2021.110054

Figure Lengend Snippet:

Article Snippet: CRT0066854 , Tocris Bioscience , Cat# 5922.

Techniques: Plasmid Preparation, Virus, Derivative Assay, Recombinant, Membrane, Protease Inhibitor, Western Blot, Lysis, Blocking Assay, Bicinchoninic Acid Protein Assay, Software

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atypical pkcs inhibitor crt0066854 hydrochloride (Tocris)

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1) Product Images from "Daphnanes diterpenes from the latex of Hura crepitans L. and their PKCζ-dependent anti-proliferative activity on colorectal cancer cells."

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