Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 4 Effects of KI forms of PKC-α, PKC-β2, PKC-δ, PKC-ε and PKC-ζ on HAA-GLUT4 translocation in rat adipocytes

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 5 Effects of WT forms of PKC-α, PKC-β1, PKC-β2, PKC-δ, PKC-ε and PKC-ζ on HAA-GLUT4 translocation in rat adipocytes

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 6 Effects of Constit forms of PKC-α, PKC-β2 and PKC-ζ on HAA-GLUT4 translocation in rat adipocytes

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 7 Effects of WT and KI forms of PKC-λ on HAA-GLUT4 translocation in rat adipocytes

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 8 Effects of combined treatment with KI forms of PKC-λ and PKC-ζ on HAA-GLUT4 translocation in rat adipocytes

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 9 Interchangeable reversing effects of WT forms of PKC-ζ and PKC-λ on HAA-GLUT4 translocation in adipocytes inhibited by KI forms of PKC-λ and PKC-ζ

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 10 Comparative effects of PMA (TPA), wortmannin, RO 31-8220, myristylated PKC-ζ/λ pseudosubstrate and GO 6976 on HAA-GLUT4 translocation (A) and 2-DOG uptake (B) in the rat adipocyte

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Biochemical Journal

Article Title: Effects of transiently expressed atypical (ζ, λ), conventional (α, β) and novel (δ, ɛ) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-ζ and C-λ

doi: 10.1042/bj3370461

Figure Lengend Snippet: Figure 11 Effects of the cell-permeant myristylated (Myr) PKC-ζ/λ pseudosubstrate (PS) on insulin-stimulated GLUT4 translocation in rat adipocytes down-regulated or non-down-regulated by prolonged phorbol ester treatment

Article Snippet: Antibodies used for Western analyses included mouse monoclonal anti-HAA antibody (Berkeley Antibody Corp.), polyclonal antisera recognizing PKC-α, PKC-β1, PKC-β2, PKCδ, the nearly identical C-termini of PKC-ζ and PKC-λ, the Nterminus of PKC-ζ and the N-terminus of PKC-λ (Santa Cruz Biotechnologies, Santa Cruz, CA, U.S.A.), polyclonal anti(PKC-ε serum) (Life Technologies, Gaithersburg, MD, U.S.A.), polyclonal anti-(Myc serum) (Upstate Biotechnologies Inc., Lake Placid, NY, U.S.A.) and mouse monoclonal anti-(PKC-λ) antibody (raised against an internal site : amino acids 397–558) (Transduction Laboratories, Lexington, KY, U.S.A.).

Techniques: Translocation Assay

Journal: Scientific Reports

Article Title: TLR4 counteracts BVRA signaling in human leukocytes via differential regulation of AMPK, mTORC1 and mTORC2

doi: 10.1038/s41598-019-43347-8

Figure Lengend Snippet: Our working model for biliverdin/BVRA and TLR4 signaling in leukocytes. Biliverdin induced responses are shown in black. Biliverdin acts on BVRA. Our data suggest that biliverdin facilitates formation of BVRA signaling complex(s) that include mTORC2. mTORC2 induces phosphorylation of two targets: PKCζ at Thr410 and Akt at Ser473. PKCζ is involved in LKB1 Ser431 and AMPKα Thr172 phosphorylation. AMPK phosphorylates TSC1/2 at Ser1387 and Raptor at Ser792. Therefore, BVRA activation may contribute to mTORC1 inhibition via two SI: phosphorylated Raptor, which inhibits mTORC1 complex formation, and TSC1/2 mediated inhibition of mTORC1 activity. Because mTORC1 is central to TLR4 signaling in leukocytes , BVRA inhibition of mTORC1 suppresses TLR4. Pharmacological inhibitors are shown in blue. Torin and PP242, but not rapamycin, inhibited BVRA-mediated protein-protein interactions. TLR4 induced response are shown in red. Once activated, TLR4 down-regulates BVRA and PKCζ expression (dashed lines). TLR4 upregulates expression of MMP9 , which then triggers AMPKα cleavage (dashed line), leading to Raptor Ser792 dephosphorylation . This enables mTORC1 activation, which then triggers an increase in HIF-1α expression and S6K1 phosphorylation at Thr389. TLR4 also triggers an increase in haptoglobin expression, thus limiting heme availability. Using these parallel mechanisms TLR4 ensures biliverdin/BVRA and mTORC2 signaling inhibition, and on the other hand, mTORC1 activation.

Article Snippet: HIF-1α (sc-10790; 1:250), AMPKα (sc-25792; 1:1000), MMP9 (sc-10737; 1:1000), TLR4 (sc-10741; 1:500), BVRA (sc-393385; 1:1000), Akt1 (sc-5928; 1:500), p-LKB1 Ser431 (sc-271924; 1:500), LKB1 (sc-32245; 1:500), p-CaMKKβ Thr286 (sc-32289; 1:750), CaMKKβ (sc-100364; 1:1000), p-mTOR Ser2481 (sc-293132; 1:1000), mTOR (sc-517464; 1:1000), p-PKCζ Thr410 (sc-271962; 1:500), PKCζ (sc-393218; 1:500), heme oxygenase 1 (HO-1) (sc-136960; 1:1000), heme oxygenase 2 (HO-2) (sc-17786; 1:1000), tuberin (TSC1/2)(sc-271314; 1:500), haptoglobin β (sc-390962; 1:500), p-NOS3 Ser1177 (sc-81510; 1:500) and NOS3 (sc-376751; 1:500) were all from Santa Cruz Biotechnology. p-Raptor Ser792 (#2083; 1:1000), Raptor (#2280; 1:200), p-p70 S6 kinase Thr389 (#9205; 1:1000), p-AMPKα Thr172 (#2535; 1:1000), p-Akt Ser473 (#4051; 1:1000), p-Akt Thr308 (#9275; 1:1000), tuberin/TSC2 (#4308; 1:1000) and p-tuberin/TSC2 Ser1387 (#5584; 1:1000) were all from Cell Signaling Technology.

Techniques: Phospho-proteomics, Activation Assay, Inhibition, Activity Assay, Protein-Protein interactions, Expressing, De-Phosphorylation Assay

Journal: Molecular medicine reports

Article Title: PKCζ, MMP‑2 and MMP‑9 expression in lung adenocarcinoma and association with a metastatic phenotype.

doi: 10.3892/mmr.2017.7634

Figure Lengend Snippet: Figure 1. Expression of PKCζ, MMP‑2 and MMP‑9 in lung adenocarcinoma, analyzed using immunohistochemistry. (A) Positive PKCζ, (B) MMP‑2 and (C) MMP‑9 staining was observed in the cytoplasm of lung adenocarcinoma sections (magnification, x100). PKCζ, protein kinase C ζ type; MMP, matrix metalloproteinase.

Article Snippet: The following commercial antibodies were used in this study: PKCζ (TA312044; 1:1,000), MMP-2 (TA806846; 1:1,000) and MMP-9 (TA353338; 1:1,000) (all from OriGene Technologies, Inc.), β-actin (4970; 1:1,000) and HRP-linked anti-rabbit IgG antibody (7074; 1:2,000) (both from Cell Signaling Technology, Inc., Danvers, MA, USA).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Molecular medicine reports

Article Title: PKCζ, MMP‑2 and MMP‑9 expression in lung adenocarcinoma and association with a metastatic phenotype.

doi: 10.3892/mmr.2017.7634

Figure Lengend Snippet: Figure 2. PKCζ siRNA‑transfected A549 cells. Following PKCζ siRNA transfection, PKCζ, MMP‑2 and MMP‑9 were immunoprecipitated and proteins were analyzed using western blotting. PKCζ, MMP‑2 and MMP‑9 expression was decreased in the siPKCζ/A549 group. PKCζ, protein kinase C ζ type; MMP, matrix metalloproteinase; siRNA, small interfering RNA; Scr, scramble.

Article Snippet: The following commercial antibodies were used in this study: PKCζ (TA312044; 1:1,000), MMP-2 (TA806846; 1:1,000) and MMP-9 (TA353338; 1:1,000) (all from OriGene Technologies, Inc.), β-actin (4970; 1:1,000) and HRP-linked anti-rabbit IgG antibody (7074; 1:2,000) (both from Cell Signaling Technology, Inc., Danvers, MA, USA).

Techniques: Transfection, Immunoprecipitation, Western Blot, Expressing, Small Interfering RNA