k ca 3 1 blockers  (R&D Systems)

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: Localization of K Ca 3.1 in ECs and SMCs of murine collateral arteries. ( a ) Representative confocal immunofluorescence images of transversal sections of collateral arteries isolated 3 h after induction of arteriogenesis. Tissue sections were stained with an antibody against K Ca 3.1 (green), together with the SMC marker αSM-actin (red), the EC marker CD31 (grey), and DAPI (blue); ( b , c ) Scatterplots showing the colocalization analysis, (left lower panel) represents pixels that have low intensity levels in both channels, green and red ( b ), or green and gray ( c ). Quadrant 4 (lower left bottom) represents pixels that are referred to as background and are not taken into consideration for colocalization analysis. Quadrant 1 represents pixels that have high green intensities and low red intensities and Quadrant 2 represents pixels that have high red intensities and low green intensities. Quadrant 3 represents pixels with high intensity levels in both green and red (b) or green and gray ( c ). These pixels are considered to be colocalized. Bright field image is also displayed. ( c ) 3D projection surface rendering is showing the localization of the K Ca 3.1 with the labelling CD 31 and αSM-actin display on the panel ( c ) right lower position. Scale bar 20 µm.

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques: Immunofluorescence, Isolation, Staining, Marker

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: Laser Doppler perfusion measurements. Line plot (left panel) along with corresponding flux images (right panel) of laser Doppler perfusion measurements. Mice were treated with solvent (control), PAP-1, or TRAM-34, respectively, and the perfusion was calculated by right to left (occlusion (occ) to sham) ratio before, immediately after, and at day 3 and 7 after the surgical procedure (left panel). Data are means ± SEM, n = 6 per group. * p < 0.05 (PAP-1 vs. control) and # p < 0.05 (TRAM-34 vs. control) from two-way ANOVA with Bonferroni’s multiple comparison test. The right panel shows representative flux images of murine paws with the tail in the center. Cold colors (blue, green) indicate low perfusion, whereas warm colors (yellow, red) indicate high perfusion (see scale).

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques:

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: BrdU incorporation and αSM-actin expression in collaterals. ( a , b ) Bar graphs represent the results of quantitative analyses of BrdU + ECs (left panels) and SMCs (right panels) in solvent (control), ( a ) PAP-1 or ( b ) TRAM-34-treated mice at day 7 after induction of arteriogenesis. Data are means ± SEM, n = 3 mice per group. * p < 0.05 from unpaired student´s t-test. The numbers of BrdU + cells in control collaterals were defined as 100%; ( c ) Representative picture of a BrdU stained collateral at day 7 after induction of arteriogenesis. Scale bar 20 µm; ( d , e ) The bar graphs represent the expression levels of αSM-actin (occlusion/sham (occ/sham)) in collateral arteries ( d ) at different time points after induction of arteriogenesis or ( e ) at 12 h after induction of arteriogenesis in control, PAP-1, or TRAM-34 treated mice. The qRT-PCR results were normalized to the expression level of the 18SrRNA. Data are means ± SEM, n > 3 per group. * p < 0.05 from unpaired student’s t-test and refers in ( d ) to occ vs. sham.

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques: BrdU Incorporation Assay, Expressing, Staining, Quantitative RT-PCR

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: Immunocytological analyses on K V 1.3 and K Ca 3.1 localization in mouse primary artery SMCs. Cells were stained with antibodies against the K V 1.3 (upper panels, green) or the K Ca 3.1 channel (middle panels, green) together with an antibody against the SMC marker αSM-actin (red) and counterstained with DAPI (blue) to show the nuclei. For negative control (lower panels) the primary antibody was omitted. Scale bar 40 µm.

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques: Staining, Marker, Negative Control

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: Proliferation assay of mouse primary artery SMCs. Mouse primary artery SMCs were cultured with 10% FCS with or without treatment of different concentrations of the K V 1.3 blocker PAP-1 or the K Ca 3.1 blocker TRAM-34. Cell proliferation was investigated by means of BrdU incorporation. Values are expressed as percentages of the positive control (+), i.e., mouse primary artery SMCs stimulated with 10% FCS. For the negative control (–), mouse primary artery SMCs cultured with 2% FCS. Data are means ± SEM, n > 6 per group. * p < 0.05 from one-way ANOVA with Bonferroni’s multiple comparison test.

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques: Proliferation Assay, Cell Culture, BrdU Incorporation Assay, Positive Control, Negative Control

Journal: Cells

Article Title: Contribution of the Potassium Channels K V 1.3 and K Ca 3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

doi: 10.3390/cells9040913

Figure Lengend Snippet: The qRT-PCR results of the expression levels of Fgfr1 , Pdgfrb, and Egr1 in vitro and during arteriogenesis in vivo. ( a , c ) Bar graphs represent the mRNA expression levels of Fgfr1, Pdgfrb, or Egr1 in vitro and ( b , d ) in vivo. In vitro mouse primary artery SMCs were cultured without (control) or with 1 μM PAP-1 or 100 nM TRAM-34, respectively. In vivo the expression level of Fgfr1 , Pdgfrb, and Egr1 were investigated 12 h after induction of arteriogenesis in collateral arteries and are expressed as occlusion (occ) to sham ratio. All qRT-PCR results were normalized to the expression level of the corresponding 18S rRNA. Data are means ± SEM, n = 3 per group. * p < 0.05 from one-way ANOVA with Bonferroni’s multiple comparison test.

Article Snippet: To block potassium channels, mice were treated either with the selective K V 1.3 channel blocker (5-(4-phenoxybutoxy)psoralen (PAP-1, 40 mg/kg/d, intraperitoneally (i.p), Sigma-Aldrich) [ ], or the selective K Ca 3.1 channel blocker TRAM-34 (120 mg/kg/d, i.p., Alomone Labs) [ ], dissolved in peanut oil, at doses previously described [ , ].

Techniques: Quantitative RT-PCR, Expressing, In Vitro, In Vivo, Cell Culture