Journal: Biochimica et biophysica acta

Article Title: The stellate vascular smooth muscle cell phenotype is induced by IL-1β via the secretion of PGE2 and subsequent cAMP-dependent protein kinase A activation.

doi: 10.1016/j.bbamcr.2015.09.019

Figure Lengend Snippet: Fig. 5. Inflammatory secretome induced by IL-1β, TNFα and IFNγ. VSMCs were treated with vehicle, IL-1β (5 ng/ml), TNFα (10 ng/ml) or IFNγ (10 ng/ml). Left panels A and B. Levels of mRNA after 48 h of stimulation, expressed as fold-changes with respect to vehicle. The diagram is based on the data shown in Table 3 (mean ± SEM of 8 independent experiments). Right panels A and B. Analysis of the cell medium 72 h after stimulation, in a membrane-based antibody array (A) and by ELISA (B). Data for the PGE2 assay and inflammatory gene mRNA levels are presented as the mean ± SEM of 6 and 8 independent experiments respectively. **, P b 0.01 versus vehicle-treated cells in Kruskal-Wallis tests followed by Dunn's multiple comparison tests. The results of the cytokine array analysis are representative of 2 independent experiments.

Article Snippet: The PGE2 receptor antagonists, L-798 106 (EP3R), SC51322 (EP1R) and ONO AE3208 (EP4R) were obtained from Tocris Bioscience, Bristol, United Kingdom.

Techniques: Membrane, Ab Array, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Biochimica et biophysica acta

Article Title: The stellate vascular smooth muscle cell phenotype is induced by IL-1β via the secretion of PGE2 and subsequent cAMP-dependent protein kinase A activation.

doi: 10.1016/j.bbamcr.2015.09.019

Figure Lengend Snippet: Fig. 6. Blocking EP4 receptors suppresses the response to conditioned medium from IL-1β-treated VSMCs (IL-1β-CM, A) and PGE2 (B). Panels A and B. Intracellular [cAMP] in naive VSMCs treated with (A) IL-1β-CM or (B) 50 nM PGE2. The various EP antagonists were added successively, as indicated in the figure. The antagonist concentrations were as follows, 280 nM for SC51322 (EP1), 6 nM for L798,106 (EP3) and 26 nM for ONO AE3208 (EP4). The black line indicates the duration of treatment. See the legend to the right panel in Fig. 3 for details. Panels C and D. Quantification of the changes in cAMP concentration induced by (C) IL-1β-CM or (D) 50 nM PGE2 in the presence or absence of the EP antagonists. n.s., non-significant, ***, P b 0.001 versus vehicle-treated cells in Kruskal–Wallis tests followed by Dunn's multiple comparison tests. Panels E and F. Phase-contrast microscopy of the changes in VSMC morphology induced by (E) IL-1β-CM or (F) 50 nM PGE2 in the presence or absence of the EP antagonists. The data and images shown are representative of 3 independent experiments.

Article Snippet: The PGE2 receptor antagonists, L-798 106 (EP3R), SC51322 (EP1R) and ONO AE3208 (EP4R) were obtained from Tocris Bioscience, Bristol, United Kingdom.

Techniques: Blocking Assay, Concentration Assay, Comparison, Microscopy

Journal: Biochimica et biophysica acta

Article Title: The stellate vascular smooth muscle cell phenotype is induced by IL-1β via the secretion of PGE2 and subsequent cAMP-dependent protein kinase A activation.

doi: 10.1016/j.bbamcr.2015.09.019

Figure Lengend Snippet: Fig. 7. Role of PKA in the development of a stellate morphology in VSMCs. Phase-contrast microscopy of naive (A) and IL-1β-transdifferentiated (B) VSMCs in the presence of vehicle, Epac (8-pCPT-O-Me-cAMP, 100 μM) and/or PKA (6BNZ-cAMP, 100 μM) activators. The concentrations used for each compound are indicated on the figure. The results shown are representative of at least 3 independent experiments. C. Numbers of copies of PKA regulatory and catalytic subunit mRNAs in VSMCs treated with vehicle or IL-1β (5 ng/ml) for 48 h. The data shown are the means ± SEM of at least 4 independent experiments. *, P b 0.05, ***, P b 0.001 versus vehicle-treated cells in Kruskal–Wallis tests followed by Dunn's multiple comparison test. D. Phase- contrast microscopy of VSMCs transfected with control siRNA (siCTRL) or siRNA directed against the PKAR1α (siR1α) or PKAR1β (siR1β) subunits. Upper and lower panels: vehicle- and PGE2−treated cells (50 nM for 1 h), respectively.

Article Snippet: The PGE2 receptor antagonists, L-798 106 (EP3R), SC51322 (EP1R) and ONO AE3208 (EP4R) were obtained from Tocris Bioscience, Bristol, United Kingdom.

Techniques: Microscopy, Comparison, Transfection, Control