Journal: The Journal of Biological Chemistry

Article Title: IL-4 up-regulates cyclooxygenase-1 expression in macrophages

doi: 10.1074/jbc.M117.785014

Figure Lengend Snippet: IL-4 stimulation increased Cox-1 protein expression. A, real-time PCR of Ptgs-1 expression in murine BMDMs stimulated with or without IL-4 and 0.1% DMSO for 20 h. B, representative Western immunoblot of Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 for 20 h. C, representative Western immunoblot of Cox-2 and Gapdh expression in murine BMDMs stimulated with or without IL-4 or LPS for 20 h (n = 1–2 biological replicates per group). D, 15d-PGJ2 production 20 h post IL-4 stimulation. E, percentage of CD206+Arg-1+F480+CD11b+ murine BMDMs stimulated with or without IL-4 relative to untreated control. F, Western immunoblots of COX-1 and GAPDH from human PBMC-derived macrophages stimulated with or without human IL-4. Unpaired two-tailed t test. **, p < 0.01; ****, p < 0.0001. n = 4–7 biological replicates per group. All experimental data are expressed as mean ± S.E.

Article Snippet: BMDMs were stimulated with or without 10 ng/ml of recombinant murine IL-4 (R&D Systems) for 20 h and analyzed for Tsc2 knock-out and Cox-1 expression by surveyor assay and Western immunoblot analysis, respectively.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Derivative Assay, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: IL-4 up-regulates cyclooxygenase-1 expression in macrophages

doi: 10.1074/jbc.M117.785014

Figure Lengend Snippet: Cox-1 is post-transcriptionally up-regulated by IL-4. A, real-time PCR on Arg-1 expression in murine BMDMs stimulated with or without IL-4 and DMSO or actinomycin D. B, representative Western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 and DMSO or actinomycin D. C, protein density of Cox-1 relative to DMSO untreated control. D, representative western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 and DMSO or cycloheximide. E, bioinformatic analysis of TOP sequences in murine and human PTGS-1 mRNA. F, polysome profile on BMDMs stimulated with or without IL-4 and torin 1 for Ptgs-1 and Rpl-13a mRNA expression, n = 1 per group. Two-way ANOVA with Fisher's LSD test. *, p < 0.05; **, p < 0.01; ****, p < 0.0001. n = 2–4 biological replicates per group. All experimental data are expressed as mean ± S.E.

Article Snippet: BMDMs were stimulated with or without 10 ng/ml of recombinant murine IL-4 (R&D Systems) for 20 h and analyzed for Tsc2 knock-out and Cox-1 expression by surveyor assay and Western immunoblot analysis, respectively.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: IL-4 up-regulates cyclooxygenase-1 expression in macrophages

doi: 10.1074/jbc.M117.785014

Figure Lengend Snippet: Cox-1 expression is controlled by the Fes/Akt/mTORC signaling pathway. A, representative Western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 and DMSO or herbimycin A. B, representative Western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 and DMSO or Akt inhibitor VIII. C, representative Western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without IL-4 and DMSO or torin 1. D, production of 15d-PGJ2 from murine BMDMs stimulated with or without IL-4 (20 h) and DMSO or torin 1. E, percentage CD206+Arg-1+F480+CD11b+ murine BMDMs stimulated with or without IL-4 or torin 1 relative to DMSO untreated control. F, Western immunoblots of COX-1 and GAPDH from human PBMC-derived macrophages stimulated with or without human IL-4 and DMSO or torin 1. G, representative Western immunoblot for Cox-1 and Gapdh expression in Cas9 murine BMDMs transduced with either Ntc or Tsc2-specific CRISPR gRNA and stimulated with or without IL-4 (n = 1 per group). Two-way ANOVA with Fisher's LSD test. *, p < 0.05; **, p < 0.01; ****, p < 0.0001. n = 3–6 biological replicates per group. All experimental data are expressed as mean ± S.E.

Article Snippet: BMDMs were stimulated with or without 10 ng/ml of recombinant murine IL-4 (R&D Systems) for 20 h and analyzed for Tsc2 knock-out and Cox-1 expression by surveyor assay and Western immunoblot analysis, respectively.

Techniques: Expressing, Western Blot, Derivative Assay, Transduction, CRISPR

Journal: The Journal of Biological Chemistry

Article Title: IL-4 up-regulates cyclooxygenase-1 expression in macrophages

doi: 10.1074/jbc.M117.785014

Figure Lengend Snippet: AMPK activation in vitro reduced Cox-1 expression and M2 macrophages. A, representative Western immunoblot for Cox-1 and Gapdh expression in murine BMDMs stimulated with or without metformin or IL-4. B, production of 15d-PGJ2 from murine BMDMs stimulated with or without metformin or IL-4 (20 h). C, percentage of CD206+Arg-1+F480+CD11b+ murine BMDMs stimulated with or without metformin or IL-4 relative to untreated control. Two-way ANOVA with Fisher's LSD test. *, p < 0.05; **, p < 0.01; ****, p < 0.0001. n = 3–7 biological replicates per group. All experimental data are expressed as mean ± S.E.

Article Snippet: BMDMs were stimulated with or without 10 ng/ml of recombinant murine IL-4 (R&D Systems) for 20 h and analyzed for Tsc2 knock-out and Cox-1 expression by surveyor assay and Western immunoblot analysis, respectively.

Techniques: Activation Assay, In Vitro, Expressing, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: IL-4 up-regulates cyclooxygenase-1 expression in macrophages

doi: 10.1074/jbc.M117.785014

Figure Lengend Snippet: Schematic illustration of IL-4-mediated post-transcriptional control of Cox-1. In addition to the previously identified Jak/Stat6 pathway that is downstream of IL-4R, our studies suggested translational control of Cox-1 expression through the IL-4R, Fes, Akt, and mTORC axis to effect M2 polarization and resolution of helminth infection. Metformin-dependent activation of AMPK inhibited mTORC to decrease M2 macrophages in addition to Cox-1 translation which reduced 15d-PGJ2 and subsequent PPARγ activation. Dotted lines represent post-transcriptional mechanisms of Cox-1 regulation downstream of mTORC that is yet to be identified.

Article Snippet: BMDMs were stimulated with or without 10 ng/ml of recombinant murine IL-4 (R&D Systems) for 20 h and analyzed for Tsc2 knock-out and Cox-1 expression by surveyor assay and Western immunoblot analysis, respectively.

Techniques: Expressing, Infection, Activation Assay

Journal: Journal of Veterinary Research

Article Title: A Pilot Study To Establish an Ovalbumin-induced Atopic Dermatitis Minipig Model

doi: 10.2478/jvetres-2021-0045

Figure Lengend Snippet: Analysis of the cytokine mRNA in AD skin as quantified by quantitative reverse transcriptase PCR A – porcine ( p ) IL-4 ; B – porcine IL-13 ; C –porcine IFN γ. Ctl – controls; DNFB – 1-fluoro-2,4-dinitrobenzene; Ova – ovalbumin. Values are mean ± SD. *p < 0.05

Article Snippet: Porcine interleukin 4 (IL-4), interferon gamma (IFNγ) and interleukin 13 (IL-13) were measured by ELISAs (cat. nos DY654 and 985; DuoSet ELISA, R&D Systems, Minneapolis, MN, USA for IL-4 and IFNγ; and cat. no. ESIL13 for IL-13; Invitrogen) according to the manufacturer’s protocols.

Techniques: Reverse Transcription

Journal: Journal of Veterinary Research

Article Title: A Pilot Study To Establish an Ovalbumin-induced Atopic Dermatitis Minipig Model

doi: 10.2478/jvetres-2021-0045

Figure Lengend Snippet: Analysis of the absolute cytokine protein level in AD skin and serum as quantified by ELISA A – porcine ( p ) IL-4; B – porcine IL-13; C –porcine IFNγ. Ctl – controls; DNFB – 1-fluoro-2,4-dinitrobenzene; Ova – ovalbumin. Values are mean ± SD. *p < 0.05

Article Snippet: Porcine interleukin 4 (IL-4), interferon gamma (IFNγ) and interleukin 13 (IL-13) were measured by ELISAs (cat. nos DY654 and 985; DuoSet ELISA, R&D Systems, Minneapolis, MN, USA for IL-4 and IFNγ; and cat. no. ESIL13 for IL-13; Invitrogen) according to the manufacturer’s protocols.

Techniques: Enzyme-linked Immunosorbent Assay