Journal: Toxins
Article Title: Veratridine Induces Vasorelaxation in Mouse Cecocolic Mesenteric Arteries
doi: 10.3390/toxins16120533
Figure Lengend Snippet: The effects of NCX antagonists on the ACh-induced Ca 2+ response in MS1 ECs. ( a ) RT-qPCR data ( left panel ) illustrated as the RNA relative level (2 −ΔCT ), showing the expression of slc8a1 , encoding NCX1. The undetectable genes ( Slc8a2 and Slc8a3 ) are not illustrated. The Western blot ( right panel ) shows the immunodetection of NCX1 in the MS1 ECs. HSC70 antibodies were used as loading controls. Protein extracts from mouse hearts served as positive controls. ( b ) An example of kinetic traces of the Fura-2 fluorescence–emission ratio, illustrating the effects of ACh (1 µM) co-injected or not co-injected with AP (100 nM), in free Na + and Ca 2+ buffers. The histograms illustrate the normalized emission ratio of Fura-2 measured after Ach injection (1 µM) in Ca 2+ -free buffer and in Na + -free buffer. These data represent the area under the curve (AUC) calculated from the kinetic traces and after normalization by ACh-induced responses at 1 µM in Hank’s Balanced Salt Solution (HBSS), used as the control. ( c , d ) The effects of NCX antagonists on the ACh-induced Ca 2+ response in MS1 ECs. The left panels show examples of kinetic traces of the Fura-2 fluorescence–emission ratio before and after the injection, at 30 s, of ACh, which led to the highest inhibition induced by the two NCX antagonists KB-R7943 (10 µM) ( c ) and SEA0400 (10 µM) ( d ). The graphs on the right panels illustrate the inhibition (% of control) induced by KB-R7943 ( c ) and SEA0400 ( d ), as a function of the ACh concentration. HBSS was used as a negative control ( d ). Data are the mean ± SEM of three independent experiments ( n = 3). Statistical significances were determined using the Mann–Whitney test ( b ) and Wilcoxon test ( c , d ) (* p < 0.05; ** p < 0.01; **** p < 0.0001; ns: non-significant).
Article Snippet: The non-specific binding sites were blocked with Tris-buffered saline solution containing 0.1% Tween and 5% BSA for 90 min. Next, the membranes were incubated with rabbit anti-NCX1 (1:500, ANX-011, Alomone Labs, Jerusalem, Israel) or rabbit anti-Pan Na V (1:200, ASC-003, Alomone Labs) primary antibodies overnight at 4 °C.
Techniques: Quantitative RT-PCR, Expressing, Western Blot, Immunodetection, Fluorescence, Injection, Control, Inhibition, Concentration Assay, Negative Control, MANN-WHITNEY