Journal: Nature Communications

Article Title: A CRISPR/Cas9-based enhancement of high-throughput single-cell transcriptomics

doi: 10.1038/s41467-025-59880-2

Figure Lengend Snippet: a Schematic figure of experimental workflow. A total of ~50,000 VSMCs were isolated from 4 donors and 2 tissue locations: coronary and pulmonary artery. b Normalized density of cells along the force directed layout (FLE) according to their artery of origin comparing 10x-V3 ( top ) with scCLEAN ( bottom ). c RNA velocity stream plots along pseudotime (CytoTRACE) mapped onto the FLE embedding. All detected genes are utilized to automatically calculate early pseudotime (black) to late pseudotime (yellow) according to the number of genes expressed. Black arrows indicate direction of transcriptional transition. d The total number of terminal states identified using CellRank comparing 10x-V3 ( left ) and scCLEAN ( right ). Number of lineages are shown. Optimal terminal states were found using Schur decomposition (gap in the real portion of eigenvalues) and then refined according to stationary distance of the coarse-grained Markov transition matrix (non-zero distance). e Only scCLEAN is shown (>1 lineage). Absorption probability of each cell belonging to each lineage and thus differentiating along pseudotime into either of the 2 terminal states, corresponding with lineage 1 ( top ) and lineage 2 ( bottom ). Yellow represents a 100% probability of that cell belonging to that lineage. f (left) Lineage absorption probability (lineage 1 = top, lineage 2 = bottom) plotted as a function of each cell’s position along the differentiation trajectory (ct pseudotime). Colors reflect tissue origin: coronary (red) or pulmonary (gray) artery. (right) Violin plots of absorption probabilities across coronary and pulmonary arteries for each lineage. g Heat matrix illustrating the relationship between the quantified absorption probability of each cell belonging to each lineage compared to the ground truth arterial identity of that cell. h Receiver operating characteristic (ROC) depicting the classification performance of identifying each artery to each lineage (both pulmonary and coronary lineage AUC = 0.99). Created in BioRender. Pandey, A. (2025) https://BioRender.com/r94y754 .

Article Snippet: Human coronary and pulmonary artery smooth muscle cells from four healthy donors were purchased from Cell Applications (San Diego, CA).

Techniques: Isolation

Journal: Biologics : Targets & Therapy

Article Title: MicroRNA-661 Suppresses Proliferation and Migration of Pulmonary Artery Smooth Muscle Cells and Endothelial Cells by Targeting CCND2

doi: 10.2147/BTT.S592879

Figure Lengend Snippet: The negative correlation between miR-661 and CCND2 in pulmonary hypertension patients. ( A ) HPASMCs were overexpressed with miR-661, followed by RNA-seq analysis. The heatmap result showed the differentially expressed genes as indicated. ( B ) qRT-PCR was used to detect the expression level of miR-661 and CCND2 in serum of patients with pulmonary hypertension and healthy volunteers, then the correlation analysis were performed. ( C ) Statistics of KEGG pathway were performed after overexpressed with miR-661 in HPASMCs, the green highlighted text indicates that the cell cycle pathway was significantly inhibited.

Article Snippet: The human embryonic Kidney 293T cells (HEK293T), human pulmonary artery smooth muscle Cells (HPASMCs) and human pulmonary artery endothelial cells (HPAECs) were obtained from the American Type Culture Collection.

Techniques: RNA Sequencing, Quantitative RT-PCR, Expressing

Journal: Physiological Reports

Article Title: The effect of mitoTEMPO on the development of hypoxia‐induced pulmonary hypertension in male mice

doi: 10.14814/phy2.70804

Figure Lengend Snippet: Effect of mitoTEMPO treatment on HIF‐1α stabilization in vitro. HIF‐1α protein levels were assessed by western blot, and steady‐state mRNA levels for lactate dehydrogenase A ( Ldha ) and pyruvate dehydrogenase kinase 1 ( Pdk1 ) by quantitative real‐time PCR in (a) CMT167 cells, (b) hPASMCs, and (c) mPASMCs, exposed to normoxia (21% O 2 ), severe hypoxia (1% O 2 ), or mild hypoxia (10% O 2 ) for 24 h and treated with triphenylphosphonium (TPP + ) (blue dots) or mitoTEMPO (MT) (red dots). Immunoblots shown are representative of three independent experiments. CMT167: mouse lung carcinoma epithelial cells; hPASMCs: human pulmonary artery smooth muscle cells; mPASMCs: mouse pulmonary artery smooth muscle cells. Densitometric analysis of HIF‐1α bands normalized to β‐Actin. Data are presented as mean ± SD. Statistical comparisons were made using two‐way ANOVA with Tukey's post hoc test ( n = 3 per group). (ns: no signal). Quantitative real‐time PCR data reflect mean ΔCt ± SD ( n = 3 per experimental group).

Article Snippet: Mouse lung carcinoma epithelial (CMT167) cells (10032302, Merck, Germany) and human PASMCs (hPASMCs) (C‐12521, PromoCell, Germany) were purchased.

Techniques: In Vitro, Western Blot, Real-time Polymerase Chain Reaction