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Inhibition of the NF-κB, p38 and JNK pathway components had an effect on the nucleic acid induced chemokine expression of NHEKs. Specific inhibiton of signaling pathway components was obtained by inhibitors (NF-κB (Bay 11−7085, 10 μM); STING (H151, 200 ng/ml); STAT-1 (Fludarabine, 10 μM); STAT-3 (Stattic, 5 μM); <t>MEK1</t> <t>(PD98059,</t> 20 μM); JNK (SP600125, 10 μM) and p38 (SB203580, 10 μM)) for 60 minutes before transfecting the cells with 0,666 μg/mL poly(I:C) or 1 ug/mL poly(dA:dT). Gene expression of fractalkine (A-C), CCL2 (D-F) and CXCL10 (G-I) was determined by the ∆∆Ct method, with GAPDH as a normalizing gene compared to the expression of the nontreated control samples. Data are presented as mean ± SD with individual datapoints, white squares representing poly(I:C), black triangles representing poly(dA:dT) treatment, n = 3. Gene expression differences were analyzed using linear mixed-effects models, p-values were adjusted using the Benjamini-Hochberg method within each solvent group-stimulus combination **** p < 0.0001, *** p < 0.001,** p < 0.01, *p < 0.05.
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biogems catalog number 1672186  (Biogems International)
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Inhibition of the NF-κB, p38 and JNK pathway components had an effect on the nucleic acid induced chemokine expression of NHEKs. Specific inhibiton of signaling pathway components was obtained by inhibitors (NF-κB (Bay 11−7085, 10 μM); STING (H151, 200 ng/ml); STAT-1 (Fludarabine, 10 μM); STAT-3 (Stattic, 5 μM); <t>MEK1</t> <t>(PD98059,</t> 20 μM); JNK (SP600125, 10 μM) and p38 (SB203580, 10 μM)) for 60 minutes before transfecting the cells with 0,666 μg/mL poly(I:C) or 1 ug/mL poly(dA:dT). Gene expression of fractalkine (A-C), CCL2 (D-F) and CXCL10 (G-I) was determined by the ∆∆Ct method, with GAPDH as a normalizing gene compared to the expression of the nontreated control samples. Data are presented as mean ± SD with individual datapoints, white squares representing poly(I:C), black triangles representing poly(dA:dT) treatment, n = 3. Gene expression differences were analyzed using linear mixed-effects models, p-values were adjusted using the Benjamini-Hochberg method within each solvent group-stimulus combination **** p < 0.0001, *** p < 0.001,** p < 0.01, *p < 0.05.
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Inhibition of the NF-κB, p38 and JNK pathway components had an effect on the nucleic acid induced chemokine expression of NHEKs. Specific inhibiton of signaling pathway components was obtained by inhibitors (NF-κB (Bay 11−7085, 10 μM); STING (H151, 200 ng/ml); STAT-1 (Fludarabine, 10 μM); STAT-3 (Stattic, 5 μM); MEK1 (PD98059, 20 μM); JNK (SP600125, 10 μM) and p38 (SB203580, 10 μM)) for 60 minutes before transfecting the cells with 0,666 μg/mL poly(I:C) or 1 ug/mL poly(dA:dT). Gene expression of fractalkine (A-C), CCL2 (D-F) and CXCL10 (G-I) was determined by the ∆∆Ct method, with GAPDH as a normalizing gene compared to the expression of the nontreated control samples. Data are presented as mean ± SD with individual datapoints, white squares representing poly(I:C), black triangles representing poly(dA:dT) treatment, n = 3. Gene expression differences were analyzed using linear mixed-effects models, p-values were adjusted using the Benjamini-Hochberg method within each solvent group-stimulus combination **** p < 0.0001, *** p < 0.001,** p < 0.01, *p < 0.05.

Journal: PLOS One

Article Title: Nucleic acid-induced chemokine expression in keratinocytes: Implications for skin inflammation

doi: 10.1371/journal.pone.0336901

Figure Lengend Snippet: Inhibition of the NF-κB, p38 and JNK pathway components had an effect on the nucleic acid induced chemokine expression of NHEKs. Specific inhibiton of signaling pathway components was obtained by inhibitors (NF-κB (Bay 11−7085, 10 μM); STING (H151, 200 ng/ml); STAT-1 (Fludarabine, 10 μM); STAT-3 (Stattic, 5 μM); MEK1 (PD98059, 20 μM); JNK (SP600125, 10 μM) and p38 (SB203580, 10 μM)) for 60 minutes before transfecting the cells with 0,666 μg/mL poly(I:C) or 1 ug/mL poly(dA:dT). Gene expression of fractalkine (A-C), CCL2 (D-F) and CXCL10 (G-I) was determined by the ∆∆Ct method, with GAPDH as a normalizing gene compared to the expression of the nontreated control samples. Data are presented as mean ± SD with individual datapoints, white squares representing poly(I:C), black triangles representing poly(dA:dT) treatment, n = 3. Gene expression differences were analyzed using linear mixed-effects models, p-values were adjusted using the Benjamini-Hochberg method within each solvent group-stimulus combination **** p < 0.0001, *** p < 0.001,** p < 0.01, *p < 0.05.

Article Snippet: For inhibition studies, cells were incubated for 60 minutes prior to poly(dA:dT)/poly(I:C) transfection with inhibitors for NF-κB (Bay 11−7085, 10 μM; MedChem Express, Monmouth Junction, NJ, USA), STING (H151, 200 ng/ml, InvivoGene) STAT-1 (Fludarabine, 10 μM; Sigma Aldrich), STAT-3 (Stattic, 5 μM; Sigma Aldrich), MEK1 (PD98059, 20 μM; Sigma Aldrich), JNK (SP600125, 10 μM; Tocris Bioscience, Bristol, UK) and p38 (SB203580, 10 μM; Tocris Bioscience).

Techniques: Inhibition, Expressing, Gene Expression, Control, Solvent