Journal: Oncotarget

Article Title: Crosstalk between insulin-like growth factor-1 and angiotensin-II in dopaminergic neurons and glial cells: role in neuroinflammation and aging

doi: 10.18632/oncotarget.9174

Figure Lengend Snippet: Western blot analysis A. - E. and real-time quantitative RT-PCR analysis F. of changes induced in the expression of angiotensin receptors (AT1 and AT2) and angiotensinogen (AGT) in primary (neuron-glia) mesencephalic cultures A. , B. , the dopaminergic cell line MES 23.5 C. , D. , the N9 microglial cell line E. and primary astroglial cultures F. Protein expression was measured relative to the α-tubulin or GAPDH band value. The results were normalized to the values for controls (100%). For RT-PCR the comparative cycle threshold values method (2 −ΔΔCt ) was used. Expression of the AGT gene was measured relative to that of the housekeeping transcripts (β-Actin). Data are means ± SEM. * p < 0.05 relative to controls. One-way ANOVA followed by Holm Sidak post-hoc test A. - D. and Student's t test E. , F.

Article Snippet: Some cultures were treated with the AT1 receptor antagonist ZD-7155 (1 μM; Tocris) or the AT2 receptor antagonist PD-123319 (1 μM; Sigma) or the NF-κB inhibitor PDTC (50μM; Sigma) for 30 minutes before treatment with AII to confirm the involvement of AT1 or AT2 receptors.

Techniques: Western Blot, Quantitative RT-PCR, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Oncotarget

Article Title: Crosstalk between insulin-like growth factor-1 and angiotensin-II in dopaminergic neurons and glial cells: role in neuroinflammation and aging

doi: 10.18632/oncotarget.9174

Figure Lengend Snippet: Western blot A. , C. , ELISA B. and Real-time quantitative RT-PCR D. of effects of IGF-1 (100 nM) on the AII-induced changes in expression of markers of the M1 cytotoxic phenotype (iNOS and TNF-α); A. , B. , a marker of the M2 repair/regenerative phenotype (ARG-1) C. , and the expression of angiotensin receptors (AT1 and AT2); D. in N9 microglial cells 24 h after treatment. Protein expression was measured relative to the GAPDH band value. The results were normalized to the values for controls (100%). TNF-α levels were expressed in pg/ml protein. For RT-PCR the comparative cycle threshold values method (2 −ΔΔCt ) was used. IGF-1 gene expression was measured relative to that of the housekeeping transcripts (β-Actin). Data are means ± SEM. * p < 0.05 relative to controls; # p < 0.05 relative to AII-treated group. One-way ANOVA followed by Holm Sidak post-hoc test A. - C. and Student's t test D. AII, angiotensin II; ARG-1, arginase-1; iNOS, inducible nitric oxide synthase; TNF-α, tumor necrosis factor alpha.

Article Snippet: Some cultures were treated with the AT1 receptor antagonist ZD-7155 (1 μM; Tocris) or the AT2 receptor antagonist PD-123319 (1 μM; Sigma) or the NF-κB inhibitor PDTC (50μM; Sigma) for 30 minutes before treatment with AII to confirm the involvement of AT1 or AT2 receptors.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Marker, Reverse Transcription Polymerase Chain Reaction, Gene Expression

Journal: Oncotarget

Article Title: Crosstalk between insulin-like growth factor-1 and angiotensin-II in dopaminergic neurons and glial cells: role in neuroinflammation and aging

doi: 10.18632/oncotarget.9174

Figure Lengend Snippet: Western blot analysis of changes induced by treatment with AII in expression of IGF-1 and IGF-1R in primary mesencephalic cultures A. , B. , E. , the dopaminergic neuron cell line MES 23.5 C. and primary mesencephalic cultures lacking microglial cells (i.e. treated with LME) D. , F. The AII-induced increase in IGF-1 and IGF-1R expression was inhibited by the AT1 receptor antagonist ZD-7155 and the AT2 agonist CG-42112A, and enhanced by the AT2 receptor antagonist PD-123319. However, treatment with AII did not induce significant changes in IGF-1 and IGF-1R in the absence of microglia C. , D. , F. Protein expression was measured relative to the α-tubulin band value. The results were normalized to the values for controls (100%). Data are means ± SEM. * p < 0.05 relative to controls; # p < 0.05 relative to AII-treated group; φ p < 0.05 relative to AII+PD group. One-way ANOVA followed by Holm Sidak post-hoc test. AII, angiotensin II; CG, AT2 agonist CG-42112A; LME, L -leucine methyl ester; PD, AT2 antagonist PD-123319; ZD, AT1 antagonist ZD-7155.

Article Snippet: Some cultures were treated with the AT1 receptor antagonist ZD-7155 (1 μM; Tocris) or the AT2 receptor antagonist PD-123319 (1 μM; Sigma) or the NF-κB inhibitor PDTC (50μM; Sigma) for 30 minutes before treatment with AII to confirm the involvement of AT1 or AT2 receptors.

Techniques: Western Blot, Expressing

Journal: Oncotarget

Article Title: Crosstalk between insulin-like growth factor-1 and angiotensin-II in dopaminergic neurons and glial cells: role in neuroinflammation and aging

doi: 10.18632/oncotarget.9174

Figure Lengend Snippet: The increase induced by AII (100 nM) in IGF-1 expression was inhibited by the AT1 receptor antagonist ZD-7155 A. and the NF-κB inhibitor PDTC C. , and enhanced by the AT2 receptor antagonist PD-123319 B. Protein expression was measured relative to the GAPDH band value. The results were normalized to the values for controls (100%). Data are means SEM. * p < 0.05 relative to controls; # p < 0.05 relative to AII-treated group; φ p < 0.05 relative to AII+PD group. One-way ANOVA followed by Holm Sidak post-hoc test AII, angiotensin II; PDTC, NF-κB inhibitor ammonium pyrrolidinedithiocarbamate; PD, AT2 antagonist PD-123319; ZD, AT1 antagonist ZD-7155.

Article Snippet: Some cultures were treated with the AT1 receptor antagonist ZD-7155 (1 μM; Tocris) or the AT2 receptor antagonist PD-123319 (1 μM; Sigma) or the NF-κB inhibitor PDTC (50μM; Sigma) for 30 minutes before treatment with AII to confirm the involvement of AT1 or AT2 receptors.

Techniques: Expressing