Journal: Journal of Behavioral Addictions

Article Title: Differential transcriptome profile underlying risky choice in a rat gambling task

doi: 10.1556/2006.2022.00068

Figure Lengend Snippet: Validation for the down-regulation of genes in risk-seeking rats. The validation experiment detected significant reductions in the expression levels for a few genes by RT-qPCR (a) and further by western blot (b), similar to the findings in RNA-seq analysis. The values are mean +S.E.M. of relative expression of molecules in the risk-seeking group compared to the risk-averse group. * P < 0.05, ** P < 0.01, risk-seeking compared to risk-averse. The number of rats are 15 and 9 for the risk-averse and risk-seeking group, respectively

Article Snippet: Antibodies used to probe the blots were as following: CaV2.3 (1:20,000; Abclonal), teneurin4 (1:1,000; Abcam), CaV1.2 (1:30,000; Alomone Labs), CaV1.3 (1:1,000; Alomone Labs), CUB and sushi domain-containing protein (1:1,000; Abclonal), and GluN2B (1:2,000, Alomone Labs) diluted in PBS-T with 5% skim milk; PKCα (1:2,000, Cell Signaling Technology), adenyl cyclase type 1 (1:2,000; Abclonal), MeCP2 (1:10,000, Cell Signaling Technology), and β-actin (1:10,000; Abcam) diluted in PBS-T with 5% BSA.

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR, Western Blot, RNA Sequencing, Analysis, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Transplantation of Neural Precursors Derived from Induced Pluripotent Cells Preserve Perineuronal Nets and Stimulate Neural Plasticity in ALS Rats

doi: 10.3390/ijms21249593

Figure Lengend Snippet: List of primary antibodies.

Article Snippet: Anti- l -type Ca 2+ CP α1C , L-type Ca 2+ CP α1C/ICC , Rabbit polyclonal , 1:200 , Alomone Labs, Jerusalem, Israel.

Techniques: Marker

Journal: Journal of Endocrinology

Article Title: CaV1.2 and CaV1.3 voltage-gated L-type Ca2+ channels in rat white fat adipocytes

doi: 10.1530/joe-19-0493

Figure Lengend Snippet: Figure 1 CaV1.1, 1.2 and 1.3 mRNA are expressed in rat adipocytes. (A) RT-CPR products (Rattus norvegicus) of voltage-dependent L-type calcium channels CaV1.1, CaV1.2, CaV1.3 and CaV1.4 alpha1 (Cacna1s, Cacna1c, Cacna1d and Cacna1f), beta2 (Cacnb1, Cacnb2, Cacnb3 and Cacnb4) and alpha2delta (Cacna2d1, Cacna2d2, Cacna2d3 and Cacna2d4) subunits for white fat adipocytes and for control: skeletal muscle for Cacna1s (CaV1.1) and whole brain for all other genes; L, DNA 50 bp ladder. PCR product sizes are in Table 1. (B) Relative expression of CaV1.1, CaV1.2 and CaV1.3 alpha-1 subunits as determined by qPCR. Data are normalized to mRNA expression of CaV1.2 in rat brain. Each point represents a different animal (n = 10), horizontal line is mean. Statistical significance is by one-way ANOVA, with Tukey’s multiple comparison test.

Article Snippet: Cells were permeablised in blocking buffer (PBS with 3% BSA and 0.5% Triton-100x) for 10 min then stained with primary anti-Calcium Channel L-type alpha 1C subunit (Cacna1C) antibody conjugated with Atto 594 (ACC-003-AG, Alomone) at 1:200 dilution for 16 h (Raifman et al. 2017).

Techniques: Control, Expressing, Comparison

Journal: Nature Communications

Article Title: Nanobody-thioesterase chimeras to specifically target protein palmitoylation

doi: 10.1038/s41467-025-56716-x

Figure Lengend Snippet: A APT2-Nanobody-F3 fusion protein recruitment to the Ca(v)1.2 complex assessed by immunoprecipitating the FLAG tagged nanobody. Experiment was repeated independently three times with similar results. B APT2 fused to Ca(v)1.2 beta subunit nanobody F3 depalmitoylates Ca(v)1.2 α1C subunit only when co-expressed with HIS-tagged beta2b subunit to facilitate nanobody recruitment to Ca(v)1.2. Data are means ± SEM from n = 5 independent experiments. Statistical comparison: unpaired two-tailed t-test. C 293 T-REx cells stably expressing α1C and transfected with β2b and α2δ1 subunits, plus either APT2-F3 or catalytically inactive (S122A) APT2-F3, were voltage-clamped at −60 mV. 500 ms voltage steps from −60 mV to +60 mV at 10 mV increments were applied at 1 Hz. Representative examples of currents from cells expressing inactive (black, left) or active (red, right) APT2 fused to nanobody F3 with barium as charge carrier are shown. D Current voltage relationship for Ca(v)1.2 mediated barium currents in the presence of active APT2-F3 (red, data are means ± SEM from n = 9 independent experiments) or catalytically inactive APT2-F3 (black, data are means ± SEM from n = 8 independent experiments). E A Boltzmann function was fitted to the I–V relationships to yield V50 and Gmax parameters for catalytically inactive APT2-F3 (black, data are means ± SEM from n = 8 independent experiments) or APT2-F3 (red, data are means ± SEM from n = 7 independent experiments). Statistical comparison: unpaired two-tailed t-test. F G–V curves were drawn by calculating G/Gmax using parameters obtained from the Boltzmann fits of I–V curves from individual cells. Data are means ± SEM from n = 8 (catalytically inactive APT2-F3, black) and n = 7 (APT2-F3, red) independent experiments. Statistical comparisons: unpaired t-test. Source data are provided as a file.

Article Snippet: This investigation used an anti-Ca(v)1.2 α1C subunit antibody raised in guinea pig obtained from Alomone Labs, and antibodies against flotillin 2 from BD Biosciences, GAPDH, FLAG and HIS tags from Merck, and GFP from Protein Tech.

Techniques: Comparison, Two Tailed Test, Stable Transfection, Expressing, Transfection, Co-Immunoprecipitation Assay, Patch Clamp

Journal: Journal of cellular physiology

Article Title: Primary Cilium Regulates CaV1.2 Expression Through Wnt Signaling

doi: 10.1002/jcp.24642

Figure Lengend Snippet: shRNA sequences

Article Snippet: Prior to experiments, cells were treated with 100 ng/ml recombinant Wnt3a (R&D Systems, Minneapolis, MN) for 3 days and serum starved for 24 h. RNAi knockdown cells shRNA lentiviral vectors specific to Cacna1c ( Origene ; pGFP-C-shLenti clone ID: TL500242) were transfected into HEK293T cells.

Techniques: shRNA, Control

Journal: Circulation: Cardiovascular Genetics

Article Title: Characterization of a Human Induced Pluripotent Stem Cell–Derived Cardiomyocyte Model for the Study of Variant Pathogenicity

doi: 10.1161/circgenetics.117.001755

Figure Lengend Snippet: Figure 4. Ion channel localization and basic electrophysiological characterization of human induced pluripotent stem cell–derived cardio- myocytes (hiPS-CMs). Shown are representative confocal images of cellular localization of (A) Kir2.1, (B) Nav1.5, and (C) Cav1.2 in hiPS under basal condition. Red fluorescent staining indicates the channels, green signal indicates α-actinin, and nuclei were stained with 4’,6-diamidino-2-phenylindole. D–F, Mean current-voltage relationship curves with typical recordings are shown in inset for total outward K+ current, n=9 (D), Na+ current, n=7 (E), and L-type Ca2+ current, n=6 (F) measured in unpaced hiPS-CMs.

Article Snippet: Antibodies used for immunofluorescence analysis Antibody Supplier Dilution Rabbit-Kir2.1 Alomone, APC026 1:200 Rabbit-Cav1.2 Alomone, ACC-022 1:200 Rabbit-Nav1.5 Alomone, ASC-005 1:200 Mouse-golgin97 (CDFX) Santa-Cruz, sc-59820 1:200 Mouse-alpha-actinin (srcomeric) Sigma Aldrich, A7811 1:2000 Alexa Fluor 555 Donkey anti-rabbit IgG Molecular probes 1:600 Alexa Fluor 488 Donkey anti-mouse IgG Molecular Probes 1:600 Supplementary table 2 (see Figure 2B).

Techniques: Stem Cell, Derivative Assay, Confocal, Staining, Immunofluorescence, Patch Clamp