Journal: Scientific reports

Article Title: ETV1 activates a rapid conduction transcriptional program in rodent and human cardiomyocytes.

doi: 10.1038/s41598-018-28239-7

Figure Lengend Snippet: Figure 2. Cardiomyocyte-specific knockout of ETV1 slows atrial and His-Purkinje system conduction. Etv1flox/

Article Snippet: P1 NRVM heart lysates were purified using Miltenyi Biotec rat neonatal cardiomyocyte isolation kit (Miltenyi Biotec, 130–105–420) according to the manufacture’s protocol.

Techniques: Knock-Out

Journal: Scientific reports

Article Title: ETV1 activates a rapid conduction transcriptional program in rodent and human cardiomyocytes.

doi: 10.1038/s41598-018-28239-7

Figure Lengend Snippet: Figure 3. Cardiomyocyte deletion of ETV1 resulted in decreased expression of fast conduction genes in atrial and His-Purkinje system (HPS) myocytes. (A) Quantitative RT-PCR of fast conduction gene RNA levels (normalized to Gapdh) comparing 10–12-week-old Etv1 WT (Etv1flox/flox) and Etv1 cKO (Etv1flox/flox, Myh6- Cre) FACS-purified ventricular, atrial, and Purkinje myocytes. Relative Nkx2–5, Gja5, and Scn5a expression displayed versus control, Etv1 WT (n = 4). (B) Immunoblot assessment of Etv1 WT and Etv1 cKO atrial tissue lysates detecting NKX2–5, Cx40, NaV1.5, and Vinculin (loading control). (C) Protein level densitometric quantification (normalized to vinculin), displayed relative to Etv1 WT (n = 5). (D) Immunofluorescence evaluation of NKX2–5, Cx40, and NaV1.5 expression in 10-week-old Etv1 WT and Etv1 cKO atria/ventricular sections. (E) Immunofluorescence evaluation of NKX2–5, Cx40, and NaV1.5 expression in 10-week-old Etv1 WT and Etv1 cKO HPS sections. Positive CNTN2 expression identified HPS cells. Nuclei were identified by DAPI (blue). LA, left atria; LV, left ventricle. Data represent mean ± SEM. *P < 0.05, 2-tailed Student’s t test. Scale bars: 50 um.

Article Snippet: P1 NRVM heart lysates were purified using Miltenyi Biotec rat neonatal cardiomyocyte isolation kit (Miltenyi Biotec, 130–105–420) according to the manufacture’s protocol.

Techniques: Expressing, Quantitative RT-PCR, Purification, Control, Western Blot, Immunofluorescence

Journal: Scientific reports

Article Title: ETV1 activates a rapid conduction transcriptional program in rodent and human cardiomyocytes.

doi: 10.1038/s41598-018-28239-7

Figure Lengend Snippet: Figure 5. ETV1 regulates the diversity of sodium channel biophysical properties between ventricular, atrial, and Purkinje myocytes. Whole-cell patch clamp data from dissociated cardiomyocytes (ventricular, right atrial, Purkinje myocytes) using 10–12 week-old Etv1 WT (Etv1flox/flox) and Etv1 cKO (Etv1flox/flox, Myh6-Cre) mice in a Cntn2-EGFP background (n = 4). (A) Comparison of sodium current–voltage (I–V) relationship. Maximum conductance was calculated to assess significant differences among experimental groups. (B) Voltage dependence of steady-state activation. Voltage at half activation (V0.5, activation) was calculated to assess significant differences among experimental groups. (C) Voltage dependence of steady-state inactivation. Voltage at half inactivation (V0.5, inactivation) was calculated to assess significant differences among experimental groups. (D) Time course of recovery from inactivation. Tau of recovery (τrecovery) was calculated to assess significant differences among experimental groups. Number of cells analyzed per cell type (ventricle, right atria, Purkinje) included in each graph legend. Patch clamp protocol diagrams are included for each endpoint. Data represent mean ± SEM. *P < 0.05, 1-way ANOVA.

Article Snippet: P1 NRVM heart lysates were purified using Miltenyi Biotec rat neonatal cardiomyocyte isolation kit (Miltenyi Biotec, 130–105–420) according to the manufacture’s protocol.

Techniques: Patch Clamp, Comparison, Activation Assay

Journal: Scientific reports

Article Title: ETV1 activates a rapid conduction transcriptional program in rodent and human cardiomyocytes.

doi: 10.1038/s41598-018-28239-7

Figure Lengend Snippet: Figure 6. ETV1-transduced neonatal rat ventricular myocytes (NRVMs) upregulates a His-Purkinje system gene signature. (A) Volcano plot of relative transcript expression from NRVMs transduced with either Ad-Etv1- EGFP or Ad-EGFP. RNA-sequencing (RNA-seq) comparison revealed a total of 9,236 differentially expressed genes (normalized counts ≥ 5, padj < 0.05). All significantly different genes (padj < 0.05) are labeled blue (downregulated) or red (enriched) and all nonsignificantly different transcripts labeled in gray. Of these there were 4,696 upregulated and 4,540 downregulated genes in Ad-Etv1-EGFP versus Ad-EGFP transduced NRVMs. (B) Functional clustering of upregulated genes in Ad-Etv1 transduced NRVMs highlighted significantly enriched ETV1-dependent cellular processes (top 20 non-redundant categories are shown). Pathways are color coded to represent genes clustered into functional classes for heat maps in C. (C) Comparative RNA-seq between 21-day-old (P21) wild-type mouse FACS-purified Purkinje cell (PC)/ventricular myocytes (VM) and Ad-Etv1-EGFP/Ad-EGFP transduced NRVMs. Heat map representation of 88 genes differentially expressed in Ad-Etv1-EGFP versus Ad-EGFP transduced NRVMs (n = 3) plotted adjacent to average fold change expression in PCs and VMs. Genes clustered into functional groups demonstrate that ETV1 regulates a PC transcriptome in neonatal cardiomyocytes.

Article Snippet: P1 NRVM heart lysates were purified using Miltenyi Biotec rat neonatal cardiomyocyte isolation kit (Miltenyi Biotec, 130–105–420) according to the manufacture’s protocol.

Techniques: Expressing, Transduction, RNA Sequencing, Comparison, Labeling, Functional Assay, Purification

Journal: Scientific reports

Article Title: ETV1 activates a rapid conduction transcriptional program in rodent and human cardiomyocytes.

doi: 10.1038/s41598-018-28239-7

Figure Lengend Snippet: Figure 8. Activation of ETV1 in human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) leads to increased expression of rapid conduction genes and sodium current. (A) Schematic representation of hiPSC-CM generation and maturation (day 0–21), transduction of Ad-Etv1-EGFP or Ad-EGFP (day 24), and timepoint for experimentation (day 38–40). (B) Quantitative RT-PCR analysis of Etv1, NKX2–5, GJA5, SCN5A, and MYL2 in hiPSC-CM transduced with either Ad-Etv1-EGFP or Ad-EGFP (n = 4). (C) Whole-cell patch clamp was performed on Ad-Etv1-EGFP (n = 12) or Ad-EGFP (n = 9) transduced hiPSC-CMs. Sodium current–voltage (I–V) relationship comparison. (D) hiPSC-CM NaV peak conductance (gNaV-peak). gNaV-peak following −120 mV to −35 mV depolarization step was measured for Ad-Etv1-EGFP (n = 12) or Ad-EGFP (n = 9) transduced hiPSC-CMs. Data represent mean ± SEM. *P < 0.05, 2-tailed Student’s t test.

Article Snippet: P1 NRVM heart lysates were purified using Miltenyi Biotec rat neonatal cardiomyocyte isolation kit (Miltenyi Biotec, 130–105–420) according to the manufacture’s protocol.

Techniques: Activation Assay, Derivative Assay, Expressing, Transduction, Quantitative RT-PCR, Patch Clamp, Comparison

Journal: Scientific reports

Article Title: Unlocking cardiac motion: assessing software and machine learning for single-cell and cardioid kinematic insights.

doi: 10.1038/s41598-024-52081-9

Figure Lengend Snippet: Figure 2. Comparison of the three open-source software programs with in-silico cardiomyocytes with 200% time-to-peak. (a) The first experiment with “grey” patterned cardiomyocyte. The plot on the left side of the panel represents the speed profile evaluated by the three software programs. The grey profile is the benchmark considered in this study (MUSCLEMOTION), the cyan profile derives from CONTRACTIONWAVE evaluation, and the orange profile is the ViKiE estimation. The histograms on the right side represent the four parameters extracted by the profiles and measured by the three methodologies. (b) The second experiment with “fog” patterned cardiomyocyte. As in panel (a), the profiles and histograms of the software evaluations are reported to address the ground-truth comparison.

Article Snippet: On the 26th day of culture, the cells were purified from all non-cardiomyocytes by MACS PSC-Derived Cardiomyocyte Isolation Kit (Miltenyi Biotec, #130-110-188) as the negative fraction to depletion antibodies.

Techniques: Comparison, Software, In Silico

Journal: American Journal of Physiology - Heart and Circulatory Physiology

Article Title: Activation of IP 3 R in atrial cardiomyocytes leads to generation of cytosolic cAMP

doi: 10.1152/ajpheart.00152.2024

Figure Lengend Snippet: Inositol trisphosphate (IP 3 ) pathway is active in human atrial induced pluripotent stem cells (iPSCs) as shown using multielectrode array. A : representative averaged multielectrode array (MEA) field action potential waveforms from control (dimethylsulfoxide, DMSO; n = 7), phenylephrine (PE; 30 μM; n = 11), and PE (1 or 30 μM) in the presence of either H89 (1 μM; n = 7) or MDL-12330A (3 μM; n = 1–10). B : effect of H89 (1 μM) and MDL-12330A (3 μM) on the beat rate of human iPSCs-derived atrial cardiomyocytes (hiPSC-ACMs) in response to PE at concentrations of 1–30 μM (DMSO, n = 7; H89, n = 7; MDL-12330A, n = 10). Data are represented as means ± SD. * P < 0.05, ** P < 0.01, and *** P < 0.001, 2-way repeated-measures ANOVA followed by Sidak’s multiple comparisons. IP 3 , inositol (1,4,5)-trisphosphate.

Article Snippet: For multielectrode array (MEA) experiments, human induced pluripotent stem cells-derived atrial cardiomyocytes (hiPSC-ACMs) (ax2518, Axol Bioscience, UK) were cultured at 5.0 × 10 5 cells/well on a 768-channel 48-well Axion Cytoview MEA 48 plate (M768-tMEA-48B, Axion Biosystems, USA) coated with Axol’s Fibronectin Coating Solution (ax0049, Axol Bioscience, UK) at 37°C in a 5% CO 2 –95% air atmosphere.

Techniques: Control, Derivative Assay

Journal: bioRxiv

Article Title: Cardiac defects in spinal muscular atrophy and the role of SMN in cardiomyocyte homeostasis

doi: 10.64898/2026.03.20.713246

Figure Lengend Snippet: KO schematic in cardiomyocytes B-D: Oxygen consumption rate (OCR) is significantly increased in siSMN-treated cardiomyocytes compared with siScramble controls and extracellular acidification rate (ECAR) is significantly decreased in siSMN-treated cardiomyocytes. n = 16 technical replicates. * p <0.05. **p < 0.01. Mitochondrial and glycolytic ATP production rates show no significant difference between siSMN and siScramble conditions (ns). E: Volcano plot of differential gene expression following SMN knockdown. F: Heatmap and hierarchical clustering of differentially expressed genes demonstrate distinct transcriptional profiles between siSMN and siScramble cardiomyocytes. G: Pathway enrichment analysis of differentially expressed genes identifies significant perturbation of multiple signaling pathways, including enrichment of PTEN signaling.

Article Snippet: Human cardiomyocytes (Axol Bioscience Limited, Cambridgeshire, England; Cat. No. ax2520; Lot No. 2520310317) were cultured and differentiated in coated plates using supplemented media from the Human iPSC-Derived Ventricular Cardiomyocyte Kit (Axol Bioscience Limited, Cambridgeshire, England).

Techniques: Gene Expression, Knockdown, Protein-Protein interactions

Journal: bioRxiv

Article Title: Cardiac defects in spinal muscular atrophy and the role of SMN in cardiomyocyte homeostasis

doi: 10.64898/2026.03.20.713246

Figure Lengend Snippet: Ingenuity pathway analysis of differentially expressed genes after SMN2 knockdown in human cardiomyocytes, showing the top 20 significantly enriched canonical pathways ranked by –log(p value). Bar color denotes predicted directionality based on z-score: black indicates negative z-score (predicted pathway inhibition), red indicates positive z-score (predicted pathway activation), and gray indicates no consistent activation pattern.

Article Snippet: Human cardiomyocytes (Axol Bioscience Limited, Cambridgeshire, England; Cat. No. ax2520; Lot No. 2520310317) were cultured and differentiated in coated plates using supplemented media from the Human iPSC-Derived Ventricular Cardiomyocyte Kit (Axol Bioscience Limited, Cambridgeshire, England).

Techniques: Knockdown, Inhibition, Activation Assay