primary antibodies against nav1 5  (Alomone Labs)

Journal: Nature Communications

Article Title: Impaired neuronal sodium channels cause intranodal conduction failure and reentrant arrhythmias in human sinoatrial node

doi: 10.1038/s41467-019-14039-8

Figure Lengend Snippet: Effects of Nav blockade on human SAN and atrial conduction

Article Snippet: Primary antibodies against Nav1.5 (custom-made in Dr Mohler’s lab), Nav1.6 (Alomone), Connexin-43(Cx43), Glyceraldehyde 3-phosphate dehydrogenase, and α-actinin (Sigma-Aldrich) were used to quantify corresponding proteins by western blotting and immunostaining , (Supplementary Table ).

Techniques:

Journal: Nature Communications

Article Title: Impaired neuronal sodium channels cause intranodal conduction failure and reentrant arrhythmias in human sinoatrial node

doi: 10.1038/s41467-019-14039-8

Figure Lengend Snippet: a Left to right: immunofluorescence image showing double staining of cNav1.5 (red) and Cx43 (green) in human heart 294050 cryosection with sinoatrial node (SAN) and right atrial (RA) regions ( n = 8) separated by white dotted line; magnification of image to show the distribution of cNav1.5 in the RA vs. the SAN; bar graph showing fluorescence signal intensity in the human SAN vs. RA. b Left to right: immunofluorescence image showing double staining of nNav1.6 (red) and Cx43 (green) in human heart cryosection with SAN and RA regions ( n = 7) separated by white dotted line; magnification of image to show the distribution of Nav1.6 in the RA vs. the SAN; bar graph showing fluorescence signal intensity in the human SAN vs. RA. c Left to right: cNav1.5 (red) and α-actinin (green; staining cardiomyocytes) dual staining; nNav1.6 (red) and α-actinin (green) dual staining confirm the cardiomyocyte-specific localization of cNav1.5 and nNav1.6. d Serial sections staining cNav1.5, nNav1.6, Cx43, and a nerve bundle labeled by anti-tyrosine hydroxylase (TH: staining sympathetic nerves) show that nNav1.6 and cNav1.5 are predominantly found in the myocytes relative to nerve bundles. All presented images a – d were collected from human heart 294050. e Left: representative immunoblotting bands for cNav1.5, α-actinin (marker of cardiomyocytes), and Cx43. Right: summary data of immunoblotting results of cNav1.5 protein distribution in the human SAN ( n = 6) and RA ( n = 6), compared with GAPDH (middle) and α-actinin (right), respectively. Cx43 connexin-43, GAPDH glyceraldehyde 3-phosphate dehydrogenase. Data were represented in mean ± SD. For immunostaining, analysis was done using lme4 and emmeans packages in R 3.4.4. Predictors included Heart (treated as random effect) and Condition (fixed effect). Pairwise tests between Condition levels were adjusted using Tukey’s method. Western blotting data analysis was done using two-sided t -test. Source data and uncropped versions of the western blot are provided as a file.

Article Snippet: Primary antibodies against Nav1.5 (custom-made in Dr Mohler’s lab), Nav1.6 (Alomone), Connexin-43(Cx43), Glyceraldehyde 3-phosphate dehydrogenase, and α-actinin (Sigma-Aldrich) were used to quantify corresponding proteins by western blotting and immunostaining , (Supplementary Table ).

Techniques: Immunofluorescence, Double Staining, Fluorescence, Staining, Labeling, Western Blot, Marker, Immunostaining

Journal: Nature Communications

Article Title: Impaired neuronal sodium channels cause intranodal conduction failure and reentrant arrhythmias in human sinoatrial node

doi: 10.1038/s41467-019-14039-8

Figure Lengend Snippet: a Geometry of 2D computer model. b Action potentials (AP) and their derivatives (d V /d t ) show different electrophysiological (EP) characteristics of SAN, SACP, and RA cells in the current model. Propagation of APs along the middle axis of the 2D SAN-atrium are displayed from the top to bottom with time at control conditions ( c ), Ado ( d ), and sodium current ( I Na ) 20% blockade ( e ). Blue and red numbers indicate the conduction time from SAN leading pacemaker through SAN and SACP, respectively. f Left: Ado plus I Na blockade reproduced prolonged SCL and SAN exit block. Right: representative APs and derivatives. Ado adenosine, RA right atria, SACP sinoatrial conduction pathway, SACT sinoatrial conduction time, SAN sinoatrial node.

Article Snippet: Primary antibodies against Nav1.5 (custom-made in Dr Mohler’s lab), Nav1.6 (Alomone), Connexin-43(Cx43), Glyceraldehyde 3-phosphate dehydrogenase, and α-actinin (Sigma-Aldrich) were used to quantify corresponding proteins by western blotting and immunostaining , (Supplementary Table ).

Techniques: Blocking Assay

Journal: Nature Communications

Article Title: Impaired neuronal sodium channels cause intranodal conduction failure and reentrant arrhythmias in human sinoatrial node

doi: 10.1038/s41467-019-14039-8

Figure Lengend Snippet: a Geometry of the 2D computer model with unexcitable fibrosis elements shown in light blue. b Action potentials (AP) and their derivatives (d V /d t ) show different electrophysiologic (EP) characteristics of SAN, SACP, and RA cells in the current model. Table results displaying combinations of adenosine dose and the percentage of sodium current ( I Na ) block in terms of cycle length, SACT, and threshold of SAN pacemaking and conduction impairment in control model ( c ) and HF model ( d ). Ado adenosine, HF heart failure, SACP sinoatrial conduction pathway, SACT sinoatrial conduction time, SAN, sinoatrial node, SCL sinus cycle length.

Article Snippet: Primary antibodies against Nav1.5 (custom-made in Dr Mohler’s lab), Nav1.6 (Alomone), Connexin-43(Cx43), Glyceraldehyde 3-phosphate dehydrogenase, and α-actinin (Sigma-Aldrich) were used to quantify corresponding proteins by western blotting and immunostaining , (Supplementary Table ).

Techniques: Blocking Assay