phospho pka substrates Search Results


ppka substrate  (Cell Signaling Technology Inc)
96
Cell Signaling Technology Inc ppka substrate
Figure 2. PDE4D promotes mTORC1 activity through Raptor Ser791 phosphorylation. (A and B) PDE4D inhibits Raptor Ser791 phosphorylation. (A) HA-tagged Raptor was coexpressed with FLAG-tagged PDE4D4 in HEK293A cells for 48 hours; cells were then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated (IP) with anti-HA antibody and immunoblotted with <t>anti-pPKA</t> <t>substrate</t> (RRXS/T) antibody. (B) Same as A but with FLAG-tagged PDE4D6. (C) PDE4D controls Raptor Ser791 phosphorylation. FLAG-tagged PDE4D (WT), FLAG-tagged PDE4D catalytically inactive mutant (D620A), and HA-tagged Raptor were overexpressed. Cells were treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-HA antibody and immunoblotted with anti-pPKA substrate antibody. (D) PDE4D depletion enhances Raptor Ser791 phosphorylation. Cells with PDE4D shRNA were stimulated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and immunoblotted for pPKA substrate. Arrow indicates PDE4D. (E) Chemical inhibition of PDE4D decreases mTORC1 activity. HEK293A cells were stimulated with rolipram, roflumilast, and GEBR- 7b for 2 hours and mTORC1 activity was analyzed. (F and G) Raptor Ser791 phosphorylation controls mTORC1 activity. (F) HEK293A cells expressing FLAG- tagged Raptor (WT) or FLAG-tagged Raptor S791A (phospho-defective) were treated with or without GEBR-7b (20 μg/mL) for 2 hours. mTORC1 activity was analyzed. (G) HA-tagged Raptor (WT) or HA-tagged Raptor S791A (phospho-defective) was overexpressed in PDE4D-depleted cells (shPDE4D). mTORC1 activity was analyzed. (H) PDE4D controls mTORC1 activity. Cells were transfected with FLAG-tagged PDE4D4 for 48 hours and stimulated with roflumi- last (50 μM) for 2 hours. mTORC1 activity was analyzed. (I and J) Pharmacologic inhibition of PDE4D enhances Raptor Ser791 phosphorylation. HEK293A cells were pretreated with either roflumilast (50 μM) (I) or GEBR-7b (20 μg/m) (J) for 1 hour and then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and Raptor Ser791 phosphorylation was assessed. Immunoblots probed for Raptor, pCREB, CREB, FLAG, HA, S6K1, and β-actin are controls. WCL, whole-cell lysates.
Ppka Substrate, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ppka substrate/product/Cell Signaling Technology Inc
Average 96 stars, based on 1 article reviews
ppka substrate - by Bioz Stars, 2026-03
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vimentin  (Cell Signaling Technology Inc)
91
Cell Signaling Technology Inc vimentin
Effect of TD-10 and TD-11 (0.005%; 48 h) on proteins involved in cell migration, apoptosis and proliferation. ( A,B) Immunostaining and immunoblotting of A549 cells treated with TEG derivatives. Downregulation of proteins involved in cell migration and EMT (mortalin, MMP2, <t>vimentin,</t> <t>VEGF,</t> β-catenin, SMAD2/3) was recorded. Downregulation of phosphorylated RB and E2F-1, Cyclin D1, and CDK4 signified moderate inhibition of cell proliferation and migration. Changes in p53, BCL2, Caspase 9 expression were mild and signified lack of apoptosis. ( C,D) Mortalin and VEGF ELISA revealed downregulation in both TD-10 and TD-11 treated cells. Statistical analysis is depicted as * p < 0.05, ** p < 0.01, *** p < 0.001.
Vimentin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/vimentin/product/Cell Signaling Technology Inc
Average 91 stars, based on 1 article reviews
vimentin - by Bioz Stars, 2026-03
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phospho-pka substrate specific antibody  (Rockland Immunochemicals)
90
Rockland Immunochemicals phospho-pka substrate specific antibody
Effect of TD-10 and TD-11 (0.005%; 48 h) on proteins involved in cell migration, apoptosis and proliferation. ( A,B) Immunostaining and immunoblotting of A549 cells treated with TEG derivatives. Downregulation of proteins involved in cell migration and EMT (mortalin, MMP2, <t>vimentin,</t> <t>VEGF,</t> β-catenin, SMAD2/3) was recorded. Downregulation of phosphorylated RB and E2F-1, Cyclin D1, and CDK4 signified moderate inhibition of cell proliferation and migration. Changes in p53, BCL2, Caspase 9 expression were mild and signified lack of apoptosis. ( C,D) Mortalin and VEGF ELISA revealed downregulation in both TD-10 and TD-11 treated cells. Statistical analysis is depicted as * p < 0.05, ** p < 0.01, *** p < 0.001.
Phospho Pka Substrate Specific Antibody, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/phospho-pka substrate specific antibody/product/Rockland Immunochemicals
Average 90 stars, based on 1 article reviews
phospho-pka substrate specific antibody - by Bioz Stars, 2026-03
90/100 stars
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Image Search Results


Figure 2. PDE4D promotes mTORC1 activity through Raptor Ser791 phosphorylation. (A and B) PDE4D inhibits Raptor Ser791 phosphorylation. (A) HA-tagged Raptor was coexpressed with FLAG-tagged PDE4D4 in HEK293A cells for 48 hours; cells were then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated (IP) with anti-HA antibody and immunoblotted with anti-pPKA substrate (RRXS/T) antibody. (B) Same as A but with FLAG-tagged PDE4D6. (C) PDE4D controls Raptor Ser791 phosphorylation. FLAG-tagged PDE4D (WT), FLAG-tagged PDE4D catalytically inactive mutant (D620A), and HA-tagged Raptor were overexpressed. Cells were treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-HA antibody and immunoblotted with anti-pPKA substrate antibody. (D) PDE4D depletion enhances Raptor Ser791 phosphorylation. Cells with PDE4D shRNA were stimulated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and immunoblotted for pPKA substrate. Arrow indicates PDE4D. (E) Chemical inhibition of PDE4D decreases mTORC1 activity. HEK293A cells were stimulated with rolipram, roflumilast, and GEBR- 7b for 2 hours and mTORC1 activity was analyzed. (F and G) Raptor Ser791 phosphorylation controls mTORC1 activity. (F) HEK293A cells expressing FLAG- tagged Raptor (WT) or FLAG-tagged Raptor S791A (phospho-defective) were treated with or without GEBR-7b (20 μg/mL) for 2 hours. mTORC1 activity was analyzed. (G) HA-tagged Raptor (WT) or HA-tagged Raptor S791A (phospho-defective) was overexpressed in PDE4D-depleted cells (shPDE4D). mTORC1 activity was analyzed. (H) PDE4D controls mTORC1 activity. Cells were transfected with FLAG-tagged PDE4D4 for 48 hours and stimulated with roflumi- last (50 μM) for 2 hours. mTORC1 activity was analyzed. (I and J) Pharmacologic inhibition of PDE4D enhances Raptor Ser791 phosphorylation. HEK293A cells were pretreated with either roflumilast (50 μM) (I) or GEBR-7b (20 μg/m) (J) for 1 hour and then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and Raptor Ser791 phosphorylation was assessed. Immunoblots probed for Raptor, pCREB, CREB, FLAG, HA, S6K1, and β-actin are controls. WCL, whole-cell lysates.

Journal: JCI insight

Article Title: Inhibition of phosphodiesterase 4D suppresses mTORC1 signaling and pancreatic cancer growth.

doi: 10.1172/jci.insight.158098

Figure Lengend Snippet: Figure 2. PDE4D promotes mTORC1 activity through Raptor Ser791 phosphorylation. (A and B) PDE4D inhibits Raptor Ser791 phosphorylation. (A) HA-tagged Raptor was coexpressed with FLAG-tagged PDE4D4 in HEK293A cells for 48 hours; cells were then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated (IP) with anti-HA antibody and immunoblotted with anti-pPKA substrate (RRXS/T) antibody. (B) Same as A but with FLAG-tagged PDE4D6. (C) PDE4D controls Raptor Ser791 phosphorylation. FLAG-tagged PDE4D (WT), FLAG-tagged PDE4D catalytically inactive mutant (D620A), and HA-tagged Raptor were overexpressed. Cells were treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-HA antibody and immunoblotted with anti-pPKA substrate antibody. (D) PDE4D depletion enhances Raptor Ser791 phosphorylation. Cells with PDE4D shRNA were stimulated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and immunoblotted for pPKA substrate. Arrow indicates PDE4D. (E) Chemical inhibition of PDE4D decreases mTORC1 activity. HEK293A cells were stimulated with rolipram, roflumilast, and GEBR- 7b for 2 hours and mTORC1 activity was analyzed. (F and G) Raptor Ser791 phosphorylation controls mTORC1 activity. (F) HEK293A cells expressing FLAG- tagged Raptor (WT) or FLAG-tagged Raptor S791A (phospho-defective) were treated with or without GEBR-7b (20 μg/mL) for 2 hours. mTORC1 activity was analyzed. (G) HA-tagged Raptor (WT) or HA-tagged Raptor S791A (phospho-defective) was overexpressed in PDE4D-depleted cells (shPDE4D). mTORC1 activity was analyzed. (H) PDE4D controls mTORC1 activity. Cells were transfected with FLAG-tagged PDE4D4 for 48 hours and stimulated with roflumi- last (50 μM) for 2 hours. mTORC1 activity was analyzed. (I and J) Pharmacologic inhibition of PDE4D enhances Raptor Ser791 phosphorylation. HEK293A cells were pretreated with either roflumilast (50 μM) (I) or GEBR-7b (20 μg/m) (J) for 1 hour and then treated with forskolin (10 μM) for 1 hour. Lysates were immunoprecipitated with anti-Raptor antibody and Raptor Ser791 phosphorylation was assessed. Immunoblots probed for Raptor, pCREB, CREB, FLAG, HA, S6K1, and β-actin are controls. WCL, whole-cell lysates.

Article Snippet: Antibodies against the following proteins were used: β-actin (catalog 3700), HA tag (catalog 3724), pULK1 (catalog 6888), ULK1 (catalog 8054), pS6K1 (catalog 9234), S6K1 (catalog 9202), p4EBP1 (catalog 2855), 4EBP1 (catalog 9452), pCREB (catalog 9198), CREB (catalog 9197), mTOR (catalog 2972), mLST8 (catalog 3274), PKA Catα (catalog 4782), PKA RIα (catalog 5675), pPKA substrate (RRXS/T; catalog 9621) (all Cell Signaling Technology); Raptor (Cell Signaling Technology, catalog 2280 and Bethyl Laboratories, catalog A300-553A), AKAP13 (Thermo Fisher Scientific, catalog PA554078), PKA RIIα (Bethyl Laboratories, catalog A301-670A-M), FLAG tag (Sigma-Aldrich, catalog F1804-50UG), PDE4A (Abcam, catalog ab200383), PDE4C (Proteintech, catalog 21754-1-AP), PDE4B (Thermo Fisher Scientific, catalog 40-1400), and PDE4D (Sigma-Aldrich, catalog ABS22; Bethyl Laboratories, catalog A303-744A; Proteintech, catalog 12918-1-AP; and Thermo Fisher Scientific, catalog PA5-21590).

Techniques: Activity Assay, Phospho-proteomics, Immunoprecipitation, Mutagenesis, shRNA, Inhibition, Expressing, Transfection, Western Blot

Effect of TD-10 and TD-11 (0.005%; 48 h) on proteins involved in cell migration, apoptosis and proliferation. ( A,B) Immunostaining and immunoblotting of A549 cells treated with TEG derivatives. Downregulation of proteins involved in cell migration and EMT (mortalin, MMP2, vimentin, VEGF, β-catenin, SMAD2/3) was recorded. Downregulation of phosphorylated RB and E2F-1, Cyclin D1, and CDK4 signified moderate inhibition of cell proliferation and migration. Changes in p53, BCL2, Caspase 9 expression were mild and signified lack of apoptosis. ( C,D) Mortalin and VEGF ELISA revealed downregulation in both TD-10 and TD-11 treated cells. Statistical analysis is depicted as * p < 0.05, ** p < 0.01, *** p < 0.001.

Journal: Frontiers in Oncology

Article Title: Identification and Functional Characterization of Anti-metastasis and Anti-angiogenic Activities of Triethylene Glycol Derivatives

doi: 10.3389/fonc.2018.00552

Figure Lengend Snippet: Effect of TD-10 and TD-11 (0.005%; 48 h) on proteins involved in cell migration, apoptosis and proliferation. ( A,B) Immunostaining and immunoblotting of A549 cells treated with TEG derivatives. Downregulation of proteins involved in cell migration and EMT (mortalin, MMP2, vimentin, VEGF, β-catenin, SMAD2/3) was recorded. Downregulation of phosphorylated RB and E2F-1, Cyclin D1, and CDK4 signified moderate inhibition of cell proliferation and migration. Changes in p53, BCL2, Caspase 9 expression were mild and signified lack of apoptosis. ( C,D) Mortalin and VEGF ELISA revealed downregulation in both TD-10 and TD-11 treated cells. Statistical analysis is depicted as * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Fixed cells were incubated with primary antibodies against β-catenin (Santa Cruz, sc-7963), BCL2 (Santa Cruz, sc-492), Caspase 9 (Santa Cruz, sc-7885), E2F-1 (Santa Cruz, sc-251), MMP2 (Santa Cruz, sc-10736), Mortalin , p53 (Santa Cruz, sc-6243), pSMAD 2/3 (Cell Signaling Technologies, 8828S), RB (Cell Signaling Technologies, 9309S), SMAD 2/3 (Santa Cruz, sc-133098), Vimentin (Santa Cruz, sc-5565), VEGF (Santa Cruz, sc-507), Cyclin D1 (Santa Cruz, sc-20044), and CDK4 (Santa Cruz, sc-260) proteins overnight, washed with PBS-PBST-PBS (5 min each), incubated with either Alexa-Fluor 488 goat anti-mouse IgG (Life Technologies, A11029) or Alexa-Fluor 594 goat anti-rabbit IgG (Life Technologies, A11037), depending upon the source of the primary antibodies, for 2 h, washed with PBS-PBST-PBS (5 min each), incubated with Hoechst 33342 stain (Invitrogen Molecular Probes, H3570) for 10 min, washed with PBST-PBS-ultrapure water (5 min each), and mounted on glass slides.

Techniques: Migration, Immunostaining, Western Blot, Inhibition, Expressing, Enzyme-linked Immunosorbent Assay