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

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Effect of hypoxia (1% O2, 24 hr) on PP2Ac activity (a, b), PP2Ac abundance (c, d), and PPP2CA mRNA expression (e, f) in HASMC and AC16 ventricular cardiomyocytes. Protein phosphatase activity is presented as a percentage of the phosphatase activity under normoxia. PP2Ac abundance and mRNA expression are normalized (β‐actin and geometric mean of GPI and GAPDH as appropriate) and expressed relative to values under normoxic conditions. Data are presented as the mean ± SEM (n = 5) and were analysed using an unpaired Student t‐test. *P < 0.05, significantly different as indicated. PP2Ac: catalytic subunit of protein phosphatase 2A; PPP2CA, gene coding PP2Ac

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Activity Assay, Expressing

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Effect of hypoxia (1% O2, 24 hr) on abundance of pPP2Ac (a, b), DPP2Ac (c, d), PME‐1 (e, f), and LCMT‐1 (g, h) in HASMC and AC16 ventricular cardiomyocytes. Representative immunoblots are inset. Normalized data are presented relative to abundance at 24 hr (mean ± SEM, n = 5) under normoxic conditions and were analysed using an unpaired Student t‐test. *P < 0.05, significantly different as indicated. DPP2Ac, demethylated protein phosphatase 2A; LCMT‐1, leucine carboxyl methyltransferase‐1; PME‐1, protein phosphatase methylesterase 1; pPP2Ac, phosphorylated catalytic subunit of protein phosphatase 2A

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Western Blot

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Effect of hypoxia (1% O2, 24 hr) on HIF‐1α abundance (a, b), total protein phosphatase activity (c, d), PP2Ac abundance (e, f), and PPP2CA mRNA expression (g, h) following HIF‐1α silencing in HASMC and AC16 ventricular cardiomyocytes. Normoxia, non‐target siRNA, and mock transfection were included as controls. HIF‐1α abundance was determined by ELISA, while total phosphatase activity, PP2Ac abundance, and PPP2CA mRNA expression were determined using a phosphatase activity assay, immunoblotting, and semi‐quantitative PCR respectively. HIF‐1α abundance is expressed in pg·ml−1. All other data are expressed relative to normoxia and where appropriate were normalized to β‐actin or geometric mean of the housekeeping genes (GPI and GAPDH). Data are presented as mean ± SEM (n = 5) and were analysed using one‐way ANOVA with Dunnett's post hoc test. *P < 0.05, significantly different as indicated. HIF‐1α, hypoxia inducible factor‐1 α; NT, non‐target siRNA; PP2Ac, catalytic subunit of protein phosphatase 2A; PPP2CA, gene coding PP2Ac; UnTx, untreated cells

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Activity Assay, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Phosphatase Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Effect of hypoxia (1% O2, 24 hr) on total phospho‐PP2Ac (a, b), demethylated PP2Ac (c, d), PME‐1 (E, F), and LCMT‐1 (g, h) abundance in HASMC and AC16 ventricular cardiomyocytes following silencing of HIF‐1α. Protein abundance was determined using immunoblotting, and all values are expressed relative to normoxia‐treated cells following normalization to β‐actin. Data are presented as mean ± SEM (n = 5) and were analysed using one‐way ANOVA with Dunnett's post hoc test. *P < 0.05, significantly different as indicated. DPP2Ac, demethylated catalytic subunit of protein phosphatase 2A; HIF‐1α, hypoxia inducible factor‐1 α; LCMT‐1, leucine carboxyl methyltransferase 1; NT, non‐target siRNA; PME‐1, protein phosphatase‐methylesterase 1; UnTx, untreated cells

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Quantitative Proteomics, Western Blot

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Effect of hypoxia (1% O2, 24 hr) on CIP2A mRNA expression (a, b) and protein abundance (c, d) in HASMC and AC16 ventricular cardiomyocytes following silencing of HIF‐1α. Protein abundance was determined using immunoblotting, and all values are expressed relative to normoxia‐treated cells following normalization to β‐actin. Data are presented as mean ± SEM (n = 5) and were analysed using one‐way ANOVA with Dunnett's post hoc test. *P < 0.05, significantly different as indicated. CIP2A, cancerous inhibitor of protein phosphatase 2A; HIF‐1α, hypoxia inducible factor‐1 α; NT, non‐target siRNA; UnTx, untreated cells

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Expressing, Quantitative Proteomics, Western Blot

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: Expression pattern of key components of the protein phosphatase system in primary human aortic endothelial (HAEC), aortic smooth muscle (HASMC), cardiac fibroblast (HCF‐av) cell lines, and AC16 ventricular cardiomyocytes: catalytic subunit expression (a, b), scaffolding subunit expression (c), regulatory subunit expression (d, e), and endogenous regulators (f). The mRNA expression profile was determined using semi‐quantitative RT‐PCR‐based SYBR green chemistry. Data are presented relative to the expression level in HAEC (mean ± SEM) and were analysed using one‐way ANOVA with post hoc analysis (Bonferroni). *P < 0.05 (n = 6). CIP2A, gene coding cancerous inhibitor of PP2A; LCMT‐1, gene coding leucine carboxyl methyltransferase 1; PPP1CA, gene coding protein phosphatase 1 catalytic subunit A; PPP2CA, gene coding protein phosphatase 2A catalytic subunit A; PPP4C, gene coding protein phosphatase 4 catalytic subunit; PPP2R1A/1B, genes coding PP2A scaffolding subunit Aα/β; PP2R2A/3A/3B/4/5A/5D, genes coding PP2A regulatory subunits; PTEN, gene coding phosphatase and tensin homologue

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Expressing, Scaffolding, Quantitative RT-PCR, SYBR Green Assay

Journal: British Journal of Pharmacology

Article Title: Hypoxia modulates protein phosphatase 2A through HIF‐1α dependent and independent mechanisms in human aortic smooth muscle cells and ventricular cardiomyocytes

doi: 10.1111/bph.14648

Figure Lengend Snippet: A schematic representation of HIF‐1α‐dependent and ‐independent regulation of the post‐translation modulation of PP2A and PP2A activity in human aortic smooth muscle cells and ventricular cardiomyocytes exposed to hypoxia (24 hr). Solid lines show experimental findings while dashed lines are inferred consequences; red and blue signify inhibition and activation respectively. CIP2A, cancerous inhibitor of PP2A; DMOG, dimethyloxalylglycine; HIF‐1α hypoxia inducible factor 1α LCMT‐1, leucine carboxyl methyltransferase 1; PDH, prolyl‐4‐hydroxylase enzyme; PP2Ac, Protein phosphatase 2A catalytic subunit A; PME‐1, protein phosphatase methylesterase‐1

Article Snippet: HASMC and HAEC were purchased from ScienCell Research Laboratories (Buckingham, UK), while ventricular fibroblasts (HCF‐av) and ventricular cardiomyocytes (AC16, Cat# SCC10; RRID:CVCL_4U18) were obtained from TCS Cellworks (Buckingham, UK) and Merck Millipore (Temecula, USA) respectively.

Techniques: Activity Assay, Inhibition, Activation Assay

Journal: JCI insight

Article Title: Mst1-mediated phosphorylation of Bcl-xL is required for myocardial reperfusion injury

doi: 10.1172/jci.insight.86217

Figure Lengend Snippet: (A) Schematic representation of the breeding strategy for the study. Red circle represents Mst1 overexpression, and blue circle represents S14A. KI, knockin. (B) Kaplan-Meier survival analysis. (C) Gross appearance of the hearts obtained from the indicated 6-month-old mice. Scale bar: 1 mm. (D) Measurement of heart-weight-to-body-weight ratio (HW/BW). (E) Representative wheat germ agglutinin staining showing cardiomyocyte cell size. Scale bar: 100 μm. (F) Quantification of relative cell size (n = 5–6). (G) Representative images showing M-mode echocardiography. Transverse scale bar: 100 ms; vertical scale bar: 5 mm. (H) Echocardiographic analyses of cardiac morphology and function (n = 7–13). LVPWd, left ventricular (LV) posterior wall dimension at diastole; LVDd, LV end-diastolic dimension; LVDs, LV end-systolic dimension; EF, ejection fraction. Data are mean ± SEM. *P < 0.05, **P < 0.001, 1-way ANOVA followed by the Newman-Keuls post-hoc analysis.

Article Snippet: Primary cultures of ventricular cardiomyocytes were prepared from 1-day-old Charles River Laboratories/Wistar Institute BR-Wistar rats (Harlan Laboratories) as described previously ( 23 ).

Techniques: Over Expression, Knock-In, Staining

Journal: iScience

Article Title: Hypoxia inhibits the cardiac I K1 current through SUMO targeting Kir2.1 activation by PIP 2

doi: 10.1016/j.isci.2022.104969

Figure Lengend Snippet:

Article Snippet: Rat ventricular cardiomyocytes from neonatal Wistar rats were purchased from Lonza Biosciences.

Techniques: Recombinant, shRNA, In Situ, Software, Microscopy