Journal: bioRxiv

Article Title: Characterization of cardiac pacemaker activity in the primate lemur Microcebus murinus

doi: 10.1101/2021.10.25.465774

Figure Lengend Snippet: A: APs recorded of SAN pacemaker myocytes from M. murinus under Tyrode’s superfusion. B: Spontaneous Ca 2+ transients recorded in SAN pacemaker myocytes under the control condition and after perfusion of Epi, ACh, ivabradine and ryanodine. C: Effect of Epi and ACh superfusion on the rate of APs (n=2 in Epi and n=2 in Ach) and Ca 2+ transients (n=2 in Epi and n=3 in Ach). D: Changes in the rate of Ca 2+ transients after perfusion of the IVA (n=4) and Ry (n=3). *p<0.05 by two-way Anova and paired T-test.

Article Snippet: Cells were then incubated for 1h at 37°C with primary antibodies against HCN4, (1:100, Alomone), sarcomeric alpha-actinin, clone EA-53 (1:100, Sigma-Aldrich), connexin 45 (1:100, H-85, Santa Cruz Biotechnology), Ca v 1.3 Ca 2+ channel polyclonal antibody (N38.8, neuromab UC Davis), cardiac troponin I (1:100, Millipore), caveolin 3 (1:100, BD Transduction Laboratories) and myogenine, (1:50, sc576 santa cruz).

Techniques: Control

Journal: The Journal of Biological Chemistry

Article Title: Regulation of Gene Transcription by Voltage-gated L-type Calcium Channel, Ca v 1.3 *

doi: 10.1074/jbc.M114.586883

Figure Lengend Snippet: 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 myocytes co-stained with anti-Cav1.3 (loop antibody, red) and α-actinin2 (green) antibodies. Scale bars are 10 μm. Right panel shows the merged image at higher magnification. d, isolated nuclei from adult atrial myocytes. Two different monoclonal antibodies for the Cav1.3 Ca2+ channel were used including: 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 the C terminus of rat Cav1.3. The monoclonal C-terminal antibody of Cav1.3 channel (C term) detects a high nuclear fluorescence in contrast to the monoclonal N-terminal antibody (N term). Nuclei were stained using DAPI (blue). Scale bar is 2 μm. The experiments were repeated independently three times.

Article Snippet: These monoclonal antibodies were obtained from the University of California Davis/National Institutes of Health NeuroMab Facility. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIGURE 3. caption a7 The C terminus of the Ca v 1.3 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 Ca v 1.3 channel (amino acids 859–875, loop antibody, upper panels ).

Techniques: Immunofluorescence, Staining, Fluorescence, Isolation