Journal: Cells

Article Title: Sympathetic Stimulation Upregulates the Ca 2+ Channel Subunit, Ca V α2δ1, via the β1 and ERK 1/2 Pathway in Neonatal Ventricular Cardiomyocytes

doi: 10.3390/cells11020188

Figure Lengend Snippet: Expression of Ca V α2δ1 ( A ), Ca V α1C ( C ), Ca V β3 ( E ), and Ca V β2 ( G ) in neonatal rat cardiomyocytes. Total proteins were extracted from neonatal rat ventricular cardiomyocytes. Proteins were separated on an 8% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane, and probed with anti-Ca V α2δ1, anti-Ca V α1C, anti-Ca V β3, anti-Ca V β2, and anti-GAPDH antibodies overnight and then incubated with HRP-conjugated goat anti-rabbit secondary antibody. Lanes were loaded with 30 μg of proteins. NE: cardiomyocytes treated with 1 µM norepinephrine. Graph showing the total protein expression of Ca V α2δ1 ( B ), Ca V α1C ( D ), Ca V β3 ( F ), and Ca V β2 ( H ) normalized to GAPDH. * p < 0.01 vs. Basal. Statistical analysis was performed using one-way ANOVA.

Article Snippet: Antibodies used were rabbit anti-Ca V α2δ1 (1:1000, Alomone ACC-015), rabbit Ca V α1C (1:1000, Alomone ACC-003), rabbit Ca V β2 (1:5000, Alomone ACC-105), rabbit Ca V β3 (1:500, Alomone ACC-008), rabbit Phospho-ERK 1/2 (Cell Signaling Technology, Danvers, MA, USA, 9101S), Total ERK 1/2 (Cell Signaling Technology, 137F5) and rabbit GAPDH (Jackson, West Grove, PA, USA, 111-035-144).

Techniques: Expressing, Incubation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Efficient stimulus-secretion coupling at ribbon synapses requires RIM-binding protein tethering of L-type Ca 2+ channels

doi: 10.1073/pnas.1702991114

Figure Lengend Snippet: Antibodies used in the current study with corresponding dilutions and origin

Article Snippet: A list of antibodies used in the current study with corresponding dilutions and origin are presented in . table ft1 table-wrap mode="anchored" t5 Table S1. caption a7 Antigen Antibody Dilution Active zone proteins RBP-1 (RIM-binding protein-1) 316003 (SySy) 1:1,000 RBP-2 (RIM-binding protein-2) 316103 (SySy) 1:1,000 RBP-2 (RIM-binding protein-2) 4193 (custom) 1:1,000 RIM 1 (RIM1 central domains) R809 (custom) 1:2,000 RIM 1/2 (1α/2α N terminus + rabphillin) U1565 (custom) 1:2,000 Liprin α3 4396 (custom) 1:5,000 ELKS 1/2aB 4790 (custom) 1:500 CASK 75–000 (Neuromab) 1:1,000 Mint1 P932 (custom) 1;1000 Veli1,2,3 T813 (custom) 1:1,000 Munc13-1 126103 (SySy) 1:1,000 Ribeye Maxeiner et al. ( 32 ) 1:1,000 Ca 2+ channels Ca 2+ v1.2-α1C voltage-gated channel ACC-003 (Alomone) 1:200 Ca 2+ v1.3-α1D voltage-gated channel ACC-005 (Alomone) 1:200 Ca 2+ v1.4-α1D voltage-gated channel Gift from Frank Schmitz 1:1,000 Ca 2+ v-α2δ1 voltage-gated channel ACC-015 (Alomone) 1:500 Ca 2+ v-α2δ2 voltage-gated channel ACC-102 (Alomone) 1:500 Ca 2+ v-α2δ3 voltage-gated channel ACC-103 (Alomone) 1:500 Ca 2+ v-α2δ4 voltage-gated channel ACC-104 (Alomone) 1:500 Ca 2+ β1 voltage-gated channel ACC-106 (Alomone) 1:250 Ca 2+ β2 voltage-gated channel ACC-105 (Alomone) 1:250 Ca 2+ β3 voltage-gated channel ACC-108 (Alomone) 1:250 Ca 2+ β4 voltage-gated channel 75–054 (Neuromab) 1:250 Exocytosis proteins Syntaxin 1 HPC-1 (custom) 1:500 SNAP 25 71.1 (custom) 1:2,000 Synaptobrevin 2 69.1 (custom) 1:2,500 Synaptotagmin 1 41.1 (custom) 1:1,000 Loading control TUJ1 T2200 (Sigma) 1:5,000 Open in a separate window Antibodies used in the current study with corresponding dilutions and origin

Techniques:

Journal: Scientific Reports

Article Title: Aberrant Splicing Promotes Proteasomal Degradation of L-type Ca V 1.2 Calcium Channels by Competitive Binding for Ca V β Subunits in Cardiac Hypertrophy

doi: 10.1038/srep35247

Figure Lengend Snippet: ( A ) Increased ratio of left ventricle to body weight in TAC mice. ( B ) Representative M-mode echocardiography images of mouse hearts before and 14 days after TAC surgery indicating progression of cardiac hypertrophy. ( C ) Increased LVAWd and LVPWd in TAC mice. ( D ) Representative gel photos for transcript screening of exons 21 + 22 inclusion level. Each lane represents a single colony expressing exons 21 + 22 or exon 21/22. ( E ) Inclusion level of exons 21 + 22 increased from 0.52% to 6.49% with the development of cardiac hypertrophy induced by pressure overload within 14 days (n = 6). ( F – H ) Expression levels of total Ca V 1.2 channels and Ca V β 2 subunits in left ventricles (n = 8). ( I , J ) Ubiquitination level of cardiac Ca V 1.2 channels in left ventricles (n = 8). Data were shown as mean ± SEM. * p < 0.05, # p < 0.01. 1-way ANOVA was performed for multiple comparisons in panel E.

Article Snippet: Subsequently, the membrane was blocked with 5% non-fat milk for 1h at room temperature and then incubated overnight at 4 °C with primary antibodies: rabbit anti-Ca V 1.2 (1:1000, ACC-003, Alomone), anti-Ca V 3.1 (1:1000, ACC-021), rabbit anti-β 2 (1:1000, ACC-105), mouse anti-ubiquitin (1:1000, 13–1600, Invitrogen), rabbit anti-HA (1:1000, 71–5500), mouse anti-TfR (1:1000, 13–6800), mouse anti-β-actin (1:5000, A1978, Sigma) or mouse anti-GAPDH (1:2000, G8795).

Techniques: Expressing