Journal: Scientific reports

Article Title: Biophysical properties of Na V 1.5 channels from atrial-like and ventricular-like cardiomyocytes derived from human induced pluripotent stem cells.

doi: 10.1038/s41598-023-47310-6

Figure Lengend Snippet: Figure 1. Characterization of specific-markers of vCMs and aCMs. (A) qPCR analysis of several cardiomyocyte genes implicated in cellular excitability (SCN5A, CACNA1C, CACNA1D, GJA1) and contraction (TNNT2, MYL2, MYL7). The analysis of exon 25 of SCN5A mRNA covered all isoforms, including the adult and neonatal isoforms. (B) Fluorescence images showing immunolabeling of cardiac TNNT2, MYL7, MYL2, ACTN1 (α-actinin), GJA1 (connexin 43), and nuclei (DAPI, cyan) (scale bar: 40 µm). Immunofluorescence images were acquired using Zeiss LSM780 confocal microscope, processed with ZEN software (Zeiss), and adapted with ImageJ software version 1.54f (NIH, Bethesda, MD, USA). (C) Western blot analysis of the expression of several excitation–contraction coupling proteins and ion channels in vCMs and aCMs. All images of cropped blot section were exposed with an optimal time to observe protein bands. All cropped blot sections were delimited by black lines. Cropped strain-free blots showing total proteins served as loading control. Original blots are presented in Suppl. Fig. S4. Top panel, middle panel, and bottom panel, respectively, refer to “Blot 1”, Blot 2” and “Blot 3″ in Suppl. Fig. S4. Cropped section areas are indicated in Suppl. Fig. S4 by red lines. Immunoblot images were adapted with ImageJ software and arranged with Microsoft Powerpoint software version microsoft 365 (Microsoft, Redmond, WA, USA).

Article Snippet: The PVDF membranes were blocked and were incubated with rabbit anti-sodium voltage-gated channel alpha subunit 5 (SCN5A) (1:200, Cat# ASC-005, Alomone Labs, RRID:AB_2040001), rabbit anti-calcium voltagegated channel subunit alpha 1 C (CACNA1C) (1:200, Cat# ACC-003, Alomone Labs, RRID:AB_2039771), mouse anti-myosin light chain 7 (MYL7) (1:400, Cat# ab68086, Abcam, RRID:AB_1140497), rabbit anti-myosin light chain 2 (MYL2) (1:2000, Cat# ab79935, Abcam, RRID:AB_1952220), mouse anti-TNNT2 (1:5000, Cat# ab10214, Abcam, RRID:AB_2206574), rabbit anti-gap junction protein alpha 1 (GJA1) (1:5000, Cat# ab11370, Abcam, RRID: AB_297976), rabbit anti-potassium voltage-gated channel subfamily A member 5 (KCNA5) (1:200, Cat# APC-150, Alomone Labs, RRID: AB_10918640), rabbit anti-ryanodine receptor 2 (RYR2) (1:1000, Cat# ARR002, Alomone Labs, RRID: AB_2040184), or rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:20,000, Cat# A300-641A, Bethyl, RRID:AB_513619).

Techniques: Fluorescence, Immunolabeling, Immunofluorescence, Microscopy, Software, Western Blot, Expressing, Control

Journal: Journal of Biological Chemistry

Article Title: Regulation of Gene Transcription by Voltage-gated L-type Calcium Channel, Cav1.3

doi: 10.1074/jbc.m114.586883

Figure Lengend Snippet: FIGURE 3. The C terminus of the Cav1.3 channel is localized in the nucleus of cardiac myocytes from wild-type mice. a, immunofluorescence confocal microscopic images of neonatal mouse cardiomyocytes stained using an antibody recognizing an epitope in the II-III cytoplasmic loop of the Cav1.3 channel (amino acids 859–875, loop antibody, upper panels). No detectable nuclear fluorescence is observed. In contrast, a high nuclear fluorescence signal is detected when an antibody that recognizes the C terminus of Cav1.3 (amino acids 1661–1990) is used (lower panels). b, isolated nuclei from adult atrial myocytes. The C-terminal antibody of the Cav1.3 channel detects a high nuclear fluorescence in contrast to the antibody directed against the II-III cytoplasmic loop. Nuclei were stained using DAPI (blue). c, adult atrial myo- cytes co-stained with anti-Cav1.3 (loop antibody, red) and -actinin2 (green) antibodies. Scale bars are 10 m. Right panel shows the merged image at higher magnification. d, isolated nuclei from adult atrial myocytes. Two dif- ferent monoclonal antibodies for the Cav1.3 Ca2 channel were used includ- ing: 1) anti-Cav1.3 Ca2 channel (NeuroMab clone L48A/9) directed against amino acids 859–875 in the N terminus of rat Cav1.3, and 2) anti-Cav1.3 Ca2 channel (NeuroMab clone N38/8) directed against amino acids 2025–2161 in

Article Snippet: 9) Monoclonal anti-Cav1.3 Ca2 channel (NeuroMab clone L48A/9) directed against amino acids 859 – 875 in the N terminus of rat Cav1.3, and 10) monoclonal anti-Cav1.3 Ca2 channel (NeuroMab clone N38/8) directed against amino acids 2025– 2161 in the C terminus of rat Cav1.3.

Techniques: Immunofluorescence, Staining, Fluorescence, Isolation, Bioprocessing