Journal: JACC: Basic to Translational Science
Article Title: Human Left Ventricle circRNA-miRNA-mRNA Network Analyses Reveal a Novel Proangiogenic Role for circNPHP1 Under Ischemic Conditions
doi: 10.1016/j.jacbts.2025.101468
Figure Lengend Snippet: CircNPHP1 and Linear NPHP1 Expression in Different Cell Types (A) Single-cell and single-nuclei RNAseq data from healthy donor hearts (human heart atlas ) of NPHP1 gene as log2 transformed counts per million (CPM). Each dot indicates the data point as 1 donor (source: human heart atlas ). (B) RNAseq: A preliminary screen was carried out in different cells (sample size = 2 each) to determine absolute levels of circNPHP1 and linear NPHP1 in human umbilical vein endothelial cells (HUVECs or HU), human cardiac microvascular endothelial cells (HCMECs or HC), human AC16 cardiomyocytes (ACs), and cardiac fibroblasts (CFs). (C) HCMEC (left) and HUVEC cells (right) were cultured in either normal conditions, hypoxia (1% oxygen), or hypoxia combined with high glucose (25 Mm D-glucose). After 48 hours, cells were harvested for quantitative real-time polymerase chain reaction for the analysis of circNPHP1 (n = 3). Fold change in RNA expression is relative to normoxia; 18S is used as housekeeping gene. Data are expressed as mean ± SEM and were assessed by 1-way analysis of variance with Dunnett's post hoc test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ns = not significant. (D) 2 μg of total RNA from HUVEC cells were digested with RNase R enzyme followed by quantitative real-time polymerase chain reaction analysis of circNPHP1 and linear NPHP1 (n = 3). Fold change in RNA expression is relative to undigested RNA; 18S is used as housekeeping gene. Data are expressed as mean ± SEM and were assessed by unpaired Student's t -test. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and ns = not significant. Abbreviations as in .
Article Snippet: Human cardiac fibroblasts (Promocell: C-12375; Lot number:491Z026.1) were cultured in fibroblast growth medium 3 (PromoCell).
Techniques: Expressing, Transformation Assay, Cell Culture, Real-time Polymerase Chain Reaction, RNA Expression