Journal: Frontiers in Pharmacology

Article Title: PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression

doi: 10.3389/fphar.2016.00139

Figure Lengend Snippet: Expression of PACAP, VIP, VIPRs, HIF-1α, HIF-2α, and EGFR in glioblastoma multiforme (GBM). (A) Representative immunoblot of PACAP and VIP precursor peptides and PAC1R, VPAC1R, and VPAC2R expression on frozen glioblastoma sample. (B) Representative immunoblot and photomicrographs of signals detected by antibodies direct against HIF-1α, HIF-2α, and EGFR in a frozen glioblastoma sample.

Article Snippet: Pituitary adenylate cyclase activating polypeptide-38 (PACAP38, 100 nM; cat no. A1439, Sigma–Aldrich), VIP, (100 nM; cat no. V3628, Sigma–Aldrich), PACAP receptor antagonist (PACAP6–38, 10 μM; cat no.3236, Tocris Biosciences, Bristol, UK) and selective VIP receptor antagonist (D-p-Cl-Phe6,Leu17-VIP, 10 μM; cat no. 3054, Tocris Biosciences) were added to U87MG cells for 24 h in normoxic or hypoxic condition.

Techniques: Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression

doi: 10.3389/fphar.2016.00139

Figure Lengend Snippet: Expression of PACAP, VIP, PAC1R, VPAC1R, and VPAC2R in glioblastoma cells under normoxic or hypoxic conditions. (A) Representative immunoblot of PACAP and VIP precursor peptides and PAC1R, VPAC1R, and VPAC2R expression on U87MG cells grown normoxia or exposed to hypoxia. (B) The bar graphs show quantitative analysis of signals obtained on immunoblots resulting from three independent experiments. Relative band densities were quantified by using ImageJ software. Protein levels are expressed as arbitrary units obtained after normalization to β-tubulin which was used as loading control. Data represent means ± SEM (* p < 0.05 or *** p < 0.001 vs. Normoxia as determined by unpaired two-tailed Student t -test).

Article Snippet: Pituitary adenylate cyclase activating polypeptide-38 (PACAP38, 100 nM; cat no. A1439, Sigma–Aldrich), VIP, (100 nM; cat no. V3628, Sigma–Aldrich), PACAP receptor antagonist (PACAP6–38, 10 μM; cat no.3236, Tocris Biosciences, Bristol, UK) and selective VIP receptor antagonist (D-p-Cl-Phe6,Leu17-VIP, 10 μM; cat no. 3054, Tocris Biosciences) were added to U87MG cells for 24 h in normoxic or hypoxic condition.

Techniques: Expressing, Western Blot, Software, Two Tailed Test

Journal: Frontiers in Pharmacology

Article Title: PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression

doi: 10.3389/fphar.2016.00139

Figure Lengend Snippet: Effect of PACAP, VIP, PI3K, and MEK1 inhibitors on U87-MG cells migration. Cells monolayer was scraped by a pipette tip and incubated with PACAP, VIP, Wortmannin or PD98059 for 24 h under normoxia or hypoxia. The wounded areas were visualized under a microscope for quantification. Migration was calculated as the average number of cells observed in five random high power wounded fields/per well in duplicate wells. In the bar graph values are expressed as percentage of control. (*** p < 0.001 vs. Vhl under normoxia; ### p < 0.001 vs. Vhl under hypoxia).

Article Snippet: Pituitary adenylate cyclase activating polypeptide-38 (PACAP38, 100 nM; cat no. A1439, Sigma–Aldrich), VIP, (100 nM; cat no. V3628, Sigma–Aldrich), PACAP receptor antagonist (PACAP6–38, 10 μM; cat no.3236, Tocris Biosciences, Bristol, UK) and selective VIP receptor antagonist (D-p-Cl-Phe6,Leu17-VIP, 10 μM; cat no. 3054, Tocris Biosciences) were added to U87MG cells for 24 h in normoxic or hypoxic condition.

Techniques: Migration, Transferring, Incubation, Microscopy

Journal: Frontiers in Pharmacology

Article Title: PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression

doi: 10.3389/fphar.2016.00139

Figure Lengend Snippet: Effect of PACAP, PACAP6-38, VIP, and VIP antagonist on expression of HIF-1α, HIF-2α, and EGFR in U87-MG cells under normoxia and hypoxia. (A) Representative immunoblots of HIF-1α, HIF-2α, and EGFR expression on U87MG cells grown normoxia or exposed to hypoxia. (B) The bar graphs show quantitative analysis of signals obtained by immunoblots resulting from three independent experiments. Relative band densities were quantified by using ImageJ software. Protein levels are expressed as arbitrary units obtained after normalization to β-tubulin which was used as loading control. Data represent means ± SEM (* p < 0.05, ** p < 0.01, or *** p < 0.001 vs. Vhl under normoxia; ### p < 0.001 vs. Vhl under hypoxia $$ p < 0.01 or $$$ p < 0.001 vs. PACAP, as determined by one-way ANOVA followed by the Tukey post hoc test).

Article Snippet: Pituitary adenylate cyclase activating polypeptide-38 (PACAP38, 100 nM; cat no. A1439, Sigma–Aldrich), VIP, (100 nM; cat no. V3628, Sigma–Aldrich), PACAP receptor antagonist (PACAP6–38, 10 μM; cat no.3236, Tocris Biosciences, Bristol, UK) and selective VIP receptor antagonist (D-p-Cl-Phe6,Leu17-VIP, 10 μM; cat no. 3054, Tocris Biosciences) were added to U87MG cells for 24 h in normoxic or hypoxic condition.

Techniques: Expressing, Western Blot, Software

Journal: Frontiers in Pharmacology

Article Title: PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression

doi: 10.3389/fphar.2016.00139

Figure Lengend Snippet: Phosphorylation of AKT and ERK1/2 in U87-MG cells under normoxia and hypoxia. (A) Representative immunoblots of Ser473-p Akt or p-Erk1/2 expression on U87MG cells treated with PACAP or VIP under normoxia or hypoxia. (B) The bar graphs show quantitative analysis of signals obtained by immunoblots resulting from three independent experiments. Relative band densities were quantified by using ImageJ software. Each signal of phosphorylated protein was normalized to total protein expression. Data are expressed as mean ± SEM (** p < 0.01, *** p < 0.001 vs. Vhl under normoxia; ### p < 0.001 vs. Vhl under hypoxia as determined by one-way ANOVA followed by the Tukey post hoc test).

Article Snippet: Pituitary adenylate cyclase activating polypeptide-38 (PACAP38, 100 nM; cat no. A1439, Sigma–Aldrich), VIP, (100 nM; cat no. V3628, Sigma–Aldrich), PACAP receptor antagonist (PACAP6–38, 10 μM; cat no.3236, Tocris Biosciences, Bristol, UK) and selective VIP receptor antagonist (D-p-Cl-Phe6,Leu17-VIP, 10 μM; cat no. 3054, Tocris Biosciences) were added to U87MG cells for 24 h in normoxic or hypoxic condition.

Techniques: Western Blot, Expressing, Software