Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Candesartan Cilexetil Attenuates Arrhythmogenicity Following Pressure Overload in Rats via the Modulation of Cardiac Electrical and Structural Remodeling and Calcium Handling Dysfunction

doi: 10.1161/JAHA.121.024285

Figure Lengend Snippet: Top in each: Representative immunoblot images of ion channel proteins. β‐actin was used as the loading control. Arrow indicates the band used for densitometric quantification. Bottom in each: Mean normalized densitometry of ion channel proteins. Bars represent mean±SD of 8, 13, 8, 12, 9, 12, 14, and 10 hearts in each group for the study of voltage‐gated potassium 4.2 channel subunit (Kv4.2), voltage‐gated potassium 4.3 channel subunit (Kv4.3), voltage‐gated potassium 1.4 channel subunit (Kv1.4), Kv‐channel interacting protein 2 (KChIP2), voltage‐gated potassium 2.1 channel subunit (Kv2.1), inwardly rectifying potassium 2.1 channel subunit (Kir2.1), voltage‐gated L‐type calcium 1.2a channel subunit (Ca V 1.2a), and voltage‐gated sodium 1.5 channel subunit (Na V 1.5), respectively. All measurements were normalized to the protein levels of β‐actin. ** P <0.01 and # P <0.001 vs Sham+Veh group; ‡ P <0.01 vs AB+Veh group by 1‐way ANOVA with a Tukey post hoc test. AB indicates aortic banding; Can, candesartan cilexetil; S, sham; and Veh, vehicle.

Article Snippet: Membranes were blocked and then incubated overnight at 4 °C with primary antibody against ryanodine receptor Ca 2+ release channel 2 (RyR2) (1:1000; ThermoFisher Scientific, Waltham, MA), Ser 2808 ‐phosphorylated RyR2 (pSer 2808 ‐RyR2) (1:5000; Badrilla, Leeds, United Kingdom), pSer 2814 ‐RyR2 (1:500; Badrilla), SERCA2a (sarco[endo]plasmic reticulum Ca 2+ ‐ATPase) (1:5000; Badrilla), phospholamban (1:2000; Badrilla), pSer 16 ‐phospholamban (1:2000; Millipore, Temecula, CA), pThr 17 ‐phospholamban (1:2000; Badrilla), Na + ‐Ca 2+ exchanger 1 (NCX1) (1:500; Santa Cruz Biotechnology, Santa Cruz, CA), voltage‐gated L‐type calcium 1.2a channel subunit (Ca V 1.2a) (1:400; Millipore), voltage‐gated sodium 1.5 channel subunit (Na V 1.5) (1:500; Alomone, Jerusalem, Israel), voltage‐gated potassium 4.2 channel subunit (Kv4.2) (1:200; Millipore), voltage‐gated potassium 4.3 channel subunit (Kv4.3) (1:200; Alomone), voltage‐gated potassium 1.4 channel subunit (Kv1.4) (1:200; Alomone), Kv‐channel interacting protein 2 (KChIP2) (1:500; ThermoFisher Scientific), voltage‐gated potassium 2.1 channel subunit (Kv2.1) (1:500; Alomone), and inwardly rectifying potassium 2.1 channel subunit (Kir2.1) (1:600; Alomone).

Techniques: Western Blot

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Notch signaling modulates the electrical behavior of cardiomyocytes

doi: 10.1152/ajpheart.00587.2016

Figure Lengend Snippet: Primers for PCR analysis

Article Snippet: Thin sections of the left ventricle (LV) were indirectly immunolabeled for the detection of GFP (Anti-GFP Antibody, catalog no. ab5450, Abcam) and KChIP2 (KCNIP2 Polyclonal Antibody, catalog no. PA5-41075, ThermoFisher Scientific).

Techniques:

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Notch signaling modulates the electrical behavior of cardiomyocytes

doi: 10.1152/ajpheart.00587.2016

Figure Lengend Snippet: Notch signaling activation reduces Kv channel-interacting protein 2 (KChIP2) expression in cardiomyocytes. A: KChIP2 (white) expression in the left ventricular (LV) myocardium of a MCM-WT mouse. Nuclei are stained by DAPI. Autofluorescence (AF) in the green channel was collected to visualize the structure of myocardial tissue. The green channel is omitted at right. B: green fluorescent protein (GFP; green) and KChIP2 (white) expression in the LV myocardium of a MCM-NICD-GFP mouse. Nuclei are stained by DAPI. The green channel is omitted at right. C: quantification of KChIP2 levels in myocytes in LV tissue of MCM-WT mice [wild type (WT), n = 2 mice] and MCM-NICD-GFP mice [Notch1 intracellular domain (NICD), n = 3 mice]. For each acquired image, the intensity of the fluorescent signal (F) from individual myocytes was normalized with respect to the signal of the myocyte presenting the highest fluorescent intensity (Fmax). Normalized data are presented as F/Fmax. *P < 0.001 vs. WT. D: quantification of relative KChIP2 levels in NICD-GFP-negative and NICD-GFP-positive myocytes in the myocardium of the MCM-NICD-GFP mice (NICD) shown in C. *P < 0.001 vs. NICD-GFP negative.

Article Snippet: Thin sections of the left ventricle (LV) were indirectly immunolabeled for the detection of GFP (Anti-GFP Antibody, catalog no. ab5450, Abcam) and KChIP2 (KCNIP2 Polyclonal Antibody, catalog no. PA5-41075, ThermoFisher Scientific).

Techniques: Activation Assay, Expressing, Staining