Journal:

Article Title: Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

doi: 10.1073/pnas.2436379100

Figure Lengend Snippet: Activation of ERK by anandamide in dormant blastocysts via CB1. (A) Localization of CB1 in dormant and activated blastocysts. The trophectoderm cell surface is decorated with CB1. The levels of CB1 are significantly lower in activated blastocysts than those in dormancy. (B) Rapid activation of ERK by anandamide (ANA) in blastocysts. Dormant blastocysts were cultured in vitro in the presence of 7 nM anandamide for the indicated times in minutes. Increased phosphorylation of ERK1/2(p-ERK1/2) and its translocation into nuclei were observed in dormant blastocyst trophectoderm cells within 5 min of their exposure to 7 nM anandamide, reaching a peak between 15 and 30 min. (C) Activation of ERK by anandamide is dose-dependent. Dormant blastocysts were cultured in vitro in the presence of 7 or 28 nM anandamide for 15 min. Anandamide at 7 nM activated ERK1/2 in dormant blastocyst trophectoderm cells, whereas it failed to do so at 28 nM. A CB1-selective antagonist SR141716A (SR1) at 7 nM or a MEK1/2 inhibitor U0126 at 1 μM inhibited the activation of ERK1/2 by 7 nM anandamide. (D) Total ERK remained unchanged. No changes in immunointensity for total ERK1/2 were observed in dormant blastocysts exposed to 7 nM anandamide or the vehicle. (E–G) Differential activation of ERK signaling by anandamide in CB mutant dormant blastocysts. CB1–/–, CB2–/–,or CB1–/– × CB2–/– dormant blastocysts were cultured in vitro in the presence of 7 nM anandamide. Activation of ERK1/2 in the presence of 7 nM anandamide for 15 min was abrogated by the CB1 antagonist SR141716A (SR1) in CB2–/– blastocyst, but not in CB1–/– or CB1–/– × CB2–/– blastocysts. Images shown depict TRITC-labeled antigens in red, Hoechst-labeled nuclei in blue, and the merge in pink. (Scale bars, 20 μm.)

Article Snippet: Rabbit polyclonal antibodies specific to CB1 (1 μg/ml, custom made), or α 1A , α 1B , and α1C (1 μg/ml, Alomone Labs, Jerusalem), or total and phospho-ERK 1 and 2 (ERK1/2) (0.5 μg/ml, New England Biolabs) were used.

Techniques: Activation Assay, Cell Culture, In Vitro, Translocation Assay, Mutagenesis, Labeling

Journal:

Article Title: Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

doi: 10.1073/pnas.2436379100

Figure Lengend Snippet: Activation of ERK by anandamide in normal day-4 blastocysts via CB1. Day-4 blastocysts were cultured in vitro in the presence of 7 or 28 nM anandamide (ANA) for 15 min. Activation of ERK1/2 at lower (7 nM) but not higher (28 nM) anandamide concentration was observed. A CB1-selective antagonist SR141716A (SR1) inhibited the accumulation of phospho-ERK1/2 (p-ERK1/2) by 7 nM anandamide. Images depict TRITC-labeled antigen in red, Hoechst-labeled nuclei in blue, and the merge in pink. (Scale bar, 20 μm.) Tr, trophectoderm; ICM, inner cell mass.

Article Snippet: Rabbit polyclonal antibodies specific to CB1 (1 μg/ml, custom made), or α 1A , α 1B , and α1C (1 μg/ml, Alomone Labs, Jerusalem), or total and phospho-ERK 1 and 2 (ERK1/2) (0.5 μg/ml, New England Biolabs) were used.

Techniques: Activation Assay, Cell Culture, In Vitro, Concentration Assay, Labeling

Journal:

Article Title: Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

doi: 10.1073/pnas.2436379100

Figure Lengend Snippet: Cannabinoid agonist CP55,940 induces activation of ERK in differentiating TS cells via CB1. (A) CB1 is expressed in TS cells. This cell line is stably transfected with the GFP gene. Images depict GFP in green, CB1 in red (TS cell surfaces), and the merge in yellow. (Scale bar, 50 μm.) (B and C) Activation of ERK in TS cells by CP55,940 (CP). TS cells were plated and expanded for 48 h. The cells were serum-starved for 5 h then exposed to different concentrations of CP for 15 min or 7 nM CP for the indicated times or to CP at 7 nM in the presence or absence of a MEK1/2 inhibitor (U0126), CB1-selective antagonist SR141716A (SR1), or CB2-selective antagonist SR144528 (SR2) for 5 min. Phosphorylation of ERK1 in differentiating TS cells was rapidly induced by 7 nM CP. U0126 or SR1, but not SR2, inhibited this activation. Quantitative analysis of ERK activation in C is expressed as percentage relative to the maximum band intensity.

Article Snippet: Rabbit polyclonal antibodies specific to CB1 (1 μg/ml, custom made), or α 1A , α 1B , and α1C (1 μg/ml, Alomone Labs, Jerusalem), or total and phospho-ERK 1 and 2 (ERK1/2) (0.5 μg/ml, New England Biolabs) were used.

Techniques: Activation Assay, Stable Transfection, Transfection

Journal:

Article Title: Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

doi: 10.1073/pnas.2436379100

Figure Lengend Snippet: Inhibition of depolarization-induced Ca2+ influx by 28 nM anandamide in dormant blastocysts. (A) Distribution of Ca2+ channel α subunits, α1B (N-type) and α1C (L-type), in dormant blastocysts. Both inner cell mass (ICM) and trophectoderm cell (Tr) are decorated with α1B and α1C subunits shown in red, Hoechst-labeled nuclei in blue, and the merge in pink. (B) Inhibition of depolarization-induced Ca2+ influx by anandamide (ANA). Ca2+ mobilization in blastocysts was visualized with Fluo-4 acetoxymethyl ester. Depolarization-induced Ca2+ influx in dormant blastocysts after exposure to 60 mM KCl was dramatically inhibited by 28 nM anandamide but not by 7 nM. The CB1-selective antagonist SR141716A (SR1) at equimolar concentration reversed this inhibition, but the CB2-selective antagonist SR144528 (SR2) was ineffective. The relative level of intracellular Ca2+ is indicated by the fluorescent intensity, which is displayed in pseudocolor according to the color bar by using lsmib. (Scale bar, 20 μm.)

Article Snippet: Rabbit polyclonal antibodies specific to CB1 (1 μg/ml, custom made), or α 1A , α 1B , and α1C (1 μg/ml, Alomone Labs, Jerusalem), or total and phospho-ERK 1 and 2 (ERK1/2) (0.5 μg/ml, New England Biolabs) were used.

Techniques: Inhibition, Labeling, Concentration Assay

Journal:

Article Title: Differential G protein-coupled cannabinoid receptor signaling by anandamide directs blastocyst activation for implantation

doi: 10.1073/pnas.2436379100

Figure Lengend Snippet: Anandamide at 7 nM confers blastocyst competency to implantation via CB1

Article Snippet: Rabbit polyclonal antibodies specific to CB1 (1 μg/ml, custom made), or α 1A , α 1B , and α1C (1 μg/ml, Alomone Labs, Jerusalem), or total and phospho-ERK 1 and 2 (ERK1/2) (0.5 μg/ml, New England Biolabs) were used.

Techniques:

Journal: Pflugers Archiv

Article Title: Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells

doi: 10.1007/s00424-010-0823-3

Figure Lengend Snippet: SNAP on TRPC5-dependent Ca 2+ -entry in HEK293 cells. a – c mTRPC5 single-cell Ca 2+ data. a , b Representative time-series graphs for [Ca 2+ ] i in vector-transfected ( b ) and mTRPC5-transfected HEK293 cells ( a ). The bath solution contained NPSS. Arrows indicate the time points when chemicals were added (SNAP, 300 µM; carbachol 100 µM; LaCl 3 100 µM; 2-APB, 75 µM). Each trace represents the mean ± SEM for ≥20 cells. c Mean ± SEM data for the [Ca 2+ ] i response 1 min after application of CCh or the maximal [Ca 2+ ] i response to La 3+ and SNAP, as illustrated in a and b . Each bar represents the mean value from seven independent experiments. d – f hTRPC5 multiwell Ca 2+ data. Shown are representative time-series graphs for [Ca 2+ ] i in response to SNAP (100 µM, 500 µM), DMSO (0.2%), and Gd 3+ (100 µM) in HEK293 cells stably expressing hTRPC5 under a tetracycline-inducible promoter with (Tet+; d ) and without tetracycline (Tet−; e ) induction. The data points represent the mean for ≥ 3 wells (SEM bars have been omitted for clarity; for full data sets see Suppl. Fig. 3). f Mean ± SEM ( n = 3) data for the maximal [Ca 2+ ] i change evoked by the applied chemicals

Article Snippet: The membrane was incubated at 4°C overnight with anti-TRPC5 or anti-TRPC6 antibody (dilution 1:200 for Alomone Lab anti-TRPC5 and anti-TRPC6, or dilution 1:500 for custom made T5C3 to peptide CKLLDSSEDVFETWGE) in PBS solution (Invitrogen) containing 0.1% Tween 20 and 5% nonfat dry milk.

Techniques: Plasmid Preparation, Transfection, Stable Transfection, Expressing

Journal: Pflugers Archiv

Article Title: Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells

doi: 10.1007/s00424-010-0823-3

Figure Lengend Snippet: DEA-NONOate on TRPC5-dependent Ca 2+ -entry. a , b Representative time-series graphs for [Ca 2+ ] i in mTRPC5-expressing HEK293 cells in response to DEA-NONOate (100 μM) ( a the data points represents the mean ± SEM for ≥20 cells in single-cell Ca 2+ measurement, final vehicle concentration of 0.1% DMSO) and in hTRPC5-expressing HEK293 cells ( b the data points represent the mean ± SEM for ≥ 3 wells in multiwell Ca 2+ measurement). c Mean ± SEM ( n ≥ 3) data for the maximal [Ca 2+ ] i change in response to DEA-NONOate as in a ( n = 8) and b ( n = 3). d Representative time-series graphs for [Ca 2+ ] i in hTRPC5-expressing HEK293 cells in response to Gd 3+ after DEA-NONOate. Carbachol (CCh), 100 µM; LaCl 3 (La 3+ ), 100 µM; GdCl 3 (Gd 3+ ), 100 µM; and DEA-NONOate (DEA), as indicated (μM); 1% DMSO in b and d

Article Snippet: The membrane was incubated at 4°C overnight with anti-TRPC5 or anti-TRPC6 antibody (dilution 1:200 for Alomone Lab anti-TRPC5 and anti-TRPC6, or dilution 1:500 for custom made T5C3 to peptide CKLLDSSEDVFETWGE) in PBS solution (Invitrogen) containing 0.1% Tween 20 and 5% nonfat dry milk.

Techniques: Expressing, Concentration Assay

Journal: Pflugers Archiv

Article Title: Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells

doi: 10.1007/s00424-010-0823-3

Figure Lengend Snippet: Ionic current through TRPC5 channels during whole-cell voltage clamp. Recordings were from HEK293 cells expressing hTRPC5. a Representative time course of whole-cell current sampled at +80 mV and −80 mV during a voltage ramp and showing bath application of 150 µM DEA-NONOate and 10 mM DTT. b , c Current–voltage relationships (I–Vs) for the experiment in a showing the initial (basal) current and the current in the presence of DEA-NONOate ( b ), and the current in the presence of DEA-NONOate and the current induced by DTT ( c ). d Mean change (▵) in inward current at −80 mV after application of 150 µM DEA-NONOate, 10 mM DTT, 500 µM SNAP (10 min) or 4 µg/ml rTRX (5 min; n ≥ 3)

Article Snippet: The membrane was incubated at 4°C overnight with anti-TRPC5 or anti-TRPC6 antibody (dilution 1:200 for Alomone Lab anti-TRPC5 and anti-TRPC6, or dilution 1:500 for custom made T5C3 to peptide CKLLDSSEDVFETWGE) in PBS solution (Invitrogen) containing 0.1% Tween 20 and 5% nonfat dry milk.

Techniques: Expressing

Journal: Pflugers Archiv

Article Title: Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells

doi: 10.1007/s00424-010-0823-3

Figure Lengend Snippet: Endogenous TRPC5 expression in BAECs and inhibitory effect of SNAP on ATP-stimulated Ca 2+ -entry involving TRPC5. a Immunocytochemical detection of TRPC5 in BAECs. The cells were stained with primary antibodies targeted to TRPC5 ( TRPC5 ; green signals ) and to platelet endothelial cell adhesion molecule 1 ( PECAM1 ; red signals ). Also shown were the merged fluorescent signals overlaid on the bright-view image ( Merged ). The control image was the merged image from sample with the omission of primary antibodies ( Control ). b Inhibitory effect of T5E3 on LaCl 3 (La 3+ , 100 µM) and carbachol (CCh, 100 µM)-induced [Ca 2+ ] i rises in mTRPC5-over-expressing HEK293 cells with or without 4 µg/ml anti-TRPC5 blocking antibody (T5E3) in the bath solution. Mean ± SEM ( n = 3) data are shown (* p < 0.05). Representative experimental traces are given in Supplementary Fig. 4. c OAG (100 µM)-induced [Ca 2+ ] i rises in TRPC3-over-expressing HEK293 cells with 4 µg/ml preimmune IgG or T5E3 in the bath solution. Mean ± SEM ( n = 3) data are shown (* p < 0.05). Representative experimental traces are given in Supplementary Fig. 4. d , e Representative time courses of [Ca 2+ ] i changes in BAECs in response to ATP (100 µM). BAECs were bathed in HBS containing 4 µg/ml T5E3 ( upper trace of e ) or preimmune IgG ( d , lower trace of e ) for 20–30 min at room temperature prior to Ca 2+ imaging. Bath solution was changed to HBS-EGTA containing preimmune IgG or T5E3 at the beginning of each recording. 300 µM SNAP ( lower trace of e ) or 0.1% DMSO ( d , upper trace of e ) was added to the bath, followed by addition of ATP to elicit store-release. After ATP-addition (8 min), the Ca 2+ influx was initiated by changing the bath solution to HBS containing ATP, DMSO/SNAP with T5E3/preimmune IgG. Each trace represents the mean ± SEM of ≥10 cells in one representative experiment. f Summary of data for all independent experiments of the type shown in d and e , showing the maximal [Ca 2+ ] i change after restoring Ca 2+ in bath ( n = 6–10; ** p < 0.01)

Article Snippet: The membrane was incubated at 4°C overnight with anti-TRPC5 or anti-TRPC6 antibody (dilution 1:200 for Alomone Lab anti-TRPC5 and anti-TRPC6, or dilution 1:500 for custom made T5C3 to peptide CKLLDSSEDVFETWGE) in PBS solution (Invitrogen) containing 0.1% Tween 20 and 5% nonfat dry milk.

Techniques: Expressing, Staining, Blocking Assay, Imaging

Journal: Pflugers Archiv

Article Title: Nitric oxide lacks direct effect on TRPC5 channels but suppresses endogenous TRPC5-containing channels in endothelial cells

doi: 10.1007/s00424-010-0823-3

Figure Lengend Snippet: Inhibitory effect of SNAP on spontaneous Ca 2+ -entry involving TRPC5 in BAECs. a , b , d , e Representative time courses of [Ca 2+ ] i changes in BAECs in response to extracellular Ca 2+ readdition. a , b Cells were incubated in HBS containing 4 µg/ml T5E3 ( upper trace of b ) or preimmune IgG ( a , lower trace of b ) for 20–30 min prior to Ca 2+ imaging. d , e Cells were transfected with empty expression vector (vector) or DN-T5 construct (DN-T5). 300 µM SNAP ( lower trace of b and e ) or 0.1% DMSO ( a and d , upper trace of b and e ) was added to the bath for 15 min, followed by changing the bath solution to HBS DMSO/SNAP with T5E3/preimmune IgG. Each trace represents the mean ± SEM of ≥5 cells in one representative experiment. c , f Summary of data for all independent experiments of the type shown in a , b (c) , d , e ( f ), showing the maximal [Ca 2+ ] i change after restoring Ca 2+ in bath. Mean ± SEM ( n = 4 for c ; n = 5–10 for f ) data are shown (* p < 0.05; ** p < 0.01)

Article Snippet: The membrane was incubated at 4°C overnight with anti-TRPC5 or anti-TRPC6 antibody (dilution 1:200 for Alomone Lab anti-TRPC5 and anti-TRPC6, or dilution 1:500 for custom made T5C3 to peptide CKLLDSSEDVFETWGE) in PBS solution (Invitrogen) containing 0.1% Tween 20 and 5% nonfat dry milk.

Techniques: Incubation, Imaging, Transfection, Expressing, Plasmid Preparation, Construct

Journal: Nature

Article Title: Mechanism of adrenergic Ca V 1.2 stimulation revealed by proximity proteomics

doi: 10.1038/s41586-020-1947-z

Figure Lengend Snippet: (a) Immunofluorescence of isolated α 1C -APEX2 and β 2B -APEX2 cardiomyocytes exposed to biotin-phenol and H 2 O 2 . Representative of 13 and 8 cardiomyocytes from 2 and 3 mice respectively. Scale bar = 5 μm. (b) Immunofluorescence of tissue sections of Langendorff-perfused α 1C -APEX2 heart. Scale bars: upper −100 μm; lower − 5 μm; lower inset− 5 μm. Representative of 10 sections from 2 mice. (c) Immunoblots of biotin-labeled proteins from α 1C -APEX2 and β 2B -APEX2 mice cardiomyocytes. In contrast to Ca V 1.2 subunits, RyR2, Jph2 and CaM, K V 1.5 channels were not detected in streptavidin-pulldown. Blots representative of 3 independent experiments. (d) Schematic of workflow for isolated cardiomyocytes. (e) Volcano plot of fold-change for relative protein quantification by TMT mass spectrometry of α 1C -APEX2 samples. Data shown are means for 5 pairs of biologically-independent samples. Non-adjusted unpaired two-tailed t-test. Rad (red dot) is reduced by 50% and PKA catalytic subunit (green dot) is increased by 50%. (f) Same as (e) except cardiomyocytes from β 2B -APEX2 mice. Data shown are means for 3 pairs of biologically independent samples. Rad is reduced by 30% and PKA catalytic subunit is increased by 68%. (g) Schematic of protein labeling and workflow for Langendorff-perfused α 1C -APEX2 mice hearts. bpm= beats per minute. (h) Same as (d) except proteins from α 1C -APEX2 whole heart samples. Data shown are means for 10 hearts, 5 without isoproterenol and 5 with isoproterenol. Rad is reduced by 36% (i) Schematic of workflow for isolated cardiomyocytes from non-transgenic (NTG) mice. (j) Same as (e) except proteins isolated from non-transgenic (NTG) mice cardiomyocytes without biotinylation or pull-down. * single peptide ESFDSQSLINNQSK. Data shown are means for 4 pairs of biologically-independent samples. Rad (red dot) in whole heart is unchanged by isoproterenol. For source gel data, see .

Article Snippet: Proteins were size-separated on SDS-PAGE, transferred to nitrocellulose membranes, and probed with anti-V5 antibody (Thermofisher, R960-25; 1:5000 dilution), a custom-made polyclonal anti-α 1C antibody (Yenzym, 1:1000 dilution) , , a custom-made polyclonal anti-β antibody (epitope: mouse residues 120-138: DSYTSRPSDSDVSLEEDRE; Yenzym, 1:1000 dilution), an anti-JPH2 antibody (Pierce, PA5-20642, lot# NG1583142; 1:1000 dilution), an anti-calmodulin antibody (Millipore Sigma 05-173; 1:1000 dilution), a custom-made anti-RyR2 antibody (1:5000 dilution) , and an anti-K V 1.5 antibody (Alomone, APC-150, Lot# APC004AN0850; 1:1000 dilution).

Techniques: Immunofluorescence, Isolation, Western Blot, Labeling, Mass Spectrometry, Two Tailed Test, Transgenic 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: 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