Journal: Experimental eye research

Article Title: The Peroxisome Proliferator-Activated Receptor-β/δ antagonist GSK0660 mitigates Retinal Cell Inflammation and Leukostasis

doi: 10.1016/j.exer.2019.107885

Figure Lengend Snippet: HRMEC were pre-treated with increasing concentrations of GSK0660 for 24hrs before stimulation with 1ng/ml TNFα for 4hrs. (A) CCL8, (B) CXCL10, and (C) CCL17 were assessed in treated HRMEC by qRT-PCR. All data are relative to the TNFα plus vehicle-treated samples. Bars represent mean ± SEM (n= 6).

Article Snippet: Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Taqman Primer Assay ID Exon boundary Amplicon Length TNFA Hs00174128_m1 3–4 80 IL1B Hs01555410_m1 3–4 91 IL6 Hs00985639_m1 2–3 66 IL8 (CXCL8) Hs00174103_m1 1–2 101 ANGPTL4 Hs01101127_m1 6–7 92 CCL8 Hs04187715_m1 1–2 67 CXCL10 Hs01124251_g1 2–3 135 CCL17 Hs00171074_m1 2–3 51 ACTB Hs99999903_m1 1–1 171 18S Hs99999901_s1 1–1 187 Open in a separate window Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.

Techniques: Quantitative RT-PCR

Journal: Experimental eye research

Article Title: The Peroxisome Proliferator-Activated Receptor-β/δ antagonist GSK0660 mitigates Retinal Cell Inflammation and Leukostasis

doi: 10.1016/j.exer.2019.107885

Figure Lengend Snippet: HRMEC monolayers were treated with vehicle, 1μM, or 10μM GSK0660 for 24hrs before stimulation with 1ng/ml TNFα. An additional group was treated with 50ng/ml of both CCL8 and CXCL10 in addition to 10μM GSK0660. PBMC were flowed over treated monolayers in a parallel plate flow chamber and adherent leukocytes were quantified. Data are shown as relative to vehicle control. Bars represent mean ± SEM (n=6).

Article Snippet: Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Taqman Primer Assay ID Exon boundary Amplicon Length TNFA Hs00174128_m1 3–4 80 IL1B Hs01555410_m1 3–4 91 IL6 Hs00985639_m1 2–3 66 IL8 (CXCL8) Hs00174103_m1 1–2 101 ANGPTL4 Hs01101127_m1 6–7 92 CCL8 Hs04187715_m1 1–2 67 CXCL10 Hs01124251_g1 2–3 135 CCL17 Hs00171074_m1 2–3 51 ACTB Hs99999903_m1 1–1 171 18S Hs99999901_s1 1–1 187 Open in a separate window Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.

Techniques: Control

Journal: Experimental eye research

Article Title: The Peroxisome Proliferator-Activated Receptor-β/δ antagonist GSK0660 mitigates Retinal Cell Inflammation and Leukostasis

doi: 10.1016/j.exer.2019.107885

Figure Lengend Snippet: HRMEC monolayers were treated with 1μg/ml anti-CCL8, 4μg/ml anti-CXCL10, or a combination of both CCL8 and CXCL10, concomitant with 1ng/ml TNFα stimulation. PBMC were flowed over treated monolayers in a parallel plate flow chamber and adherent leukocytes were quantified. Data are shown as relative to vehicle control. Bars represent mean ± SEM (n=6).

Article Snippet: Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Taqman Primer Assay ID Exon boundary Amplicon Length TNFA Hs00174128_m1 3–4 80 IL1B Hs01555410_m1 3–4 91 IL6 Hs00985639_m1 2–3 66 IL8 (CXCL8) Hs00174103_m1 1–2 101 ANGPTL4 Hs01101127_m1 6–7 92 CCL8 Hs04187715_m1 1–2 67 CXCL10 Hs01124251_g1 2–3 135 CCL17 Hs00171074_m1 2–3 51 ACTB Hs99999903_m1 1–1 171 18S Hs99999901_s1 1–1 187 Open in a separate window Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.

Techniques: Control

Journal: Experimental eye research

Article Title: The Peroxisome Proliferator-Activated Receptor-β/δ antagonist GSK0660 mitigates Retinal Cell Inflammation and Leukostasis

doi: 10.1016/j.exer.2019.107885

Figure Lengend Snippet: (A) Mice were injected with vehicle, 1μM GW0742, 50ng/ml TNFα vehicle, or 50ng/ml TNFα with 1μM GSK0660. (B) Mice were injected with vehicle or 50ng/ml recombinant CCL8 with 50ng/ml recombinant CXCL10. Six hrs after intravitreal injection, mice were perfused with FITC-conjugated concanavalin-A to label adherent leukocytes. Bars represent mean ± SEM (A: vehicle n=13; GW0742 n=5; TNFα vehicle n=4; TNFα + GSK0660 n=8; B: vehicle n=5; CCL8/CXCL10 n=5).

Article Snippet: Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Taqman Primer Assay ID Exon boundary Amplicon Length TNFA Hs00174128_m1 3–4 80 IL1B Hs01555410_m1 3–4 91 IL6 Hs00985639_m1 2–3 66 IL8 (CXCL8) Hs00174103_m1 1–2 101 ANGPTL4 Hs01101127_m1 6–7 92 CCL8 Hs04187715_m1 1–2 67 CXCL10 Hs01124251_g1 2–3 135 CCL17 Hs00171074_m1 2–3 51 ACTB Hs99999903_m1 1–1 171 18S Hs99999901_s1 1–1 187 Open in a separate window Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.

Techniques: Injection, Recombinant

Journal: Experimental eye research

Article Title: The Peroxisome Proliferator-Activated Receptor-β/δ antagonist GSK0660 mitigates Retinal Cell Inflammation and Leukostasis

doi: 10.1016/j.exer.2019.107885

Figure Lengend Snippet: Palmitic acid stimulates the release of inflammatory cytokines (TNFα, IL-1β, IL-6, IL-8) and a pro-angiogenic factor (Angptl4). GSK0660 reduces the stimulation of these factors in PPARβ/δ-independent and -dependent manners, respectively. TNFα can then act in a paracrine manner to stimulate CCL8 and CXCL10 production by endothelial cells. GSK0660 blocks the production of these chemokines in a PPARβ/δ-dependent manner, thereby limiting leukostasis.

Article Snippet: Data were analyzed using the comparative Ct method and Ct values were normalized to ACTB or 18S levels. table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene Taqman Primer Assay ID Exon boundary Amplicon Length TNFA Hs00174128_m1 3–4 80 IL1B Hs01555410_m1 3–4 91 IL6 Hs00985639_m1 2–3 66 IL8 (CXCL8) Hs00174103_m1 1–2 101 ANGPTL4 Hs01101127_m1 6–7 92 CCL8 Hs04187715_m1 1–2 67 CXCL10 Hs01124251_g1 2–3 135 CCL17 Hs00171074_m1 2–3 51 ACTB Hs99999903_m1 1–1 171 18S Hs99999901_s1 1–1 187 Open in a separate window Primer IDs, exons boundary, and amplicon length of Taqman primers used in this study.

Techniques: