cells  (PromoCell)

Journal: Journal of Extracellular Vesicles

Article Title: The Fatty Acid Transporter CD36 Mediates Uptake, Biodistribution, and Cardioprotection by Small Extracellular Vesicles From HEK293 Cells

doi: 10.1002/jev2.70254

Figure Lengend Snippet: Dose‐response and time‐response uptake of HEK293‐sEVs in human endothelial cells and cardiomyocytes. (A–C) Human coronary artery and coronary microvascular endothelial cells (HCAEC and HCMEC respectively) and human umbilical vein endothelial cells (HUVEC) were treated for 4 h with increasing doses of HEK293‐sEVs. A linear dose‐response was revealed in all endothelial cells, as a 10‐fold increase of sEVs dose results in a 10‐fold increase of cellular uptake (Nluc Nanoluciferase activity). (D–F) sEVs rapidly internalise endothelial cells ((D) HCAEC, (E) HCMEC, and (F) HUVEC) within the first hour of incubation reaching the highest intracellular localisation after 4 h, corresponding to higher Nluc activity. A reduction in luciferase activity after 24 h is observed in all endothelial cells, reflective of the reduced amount of sEVs intracellularly. (G) Uptake of HEK293‐sEVs in adult rat cardiomyocytes is slow and time‐dependent, and high Nluc activity is recorded only after 24 h of sEVs incubation. Data shown as mean ± SEM ( n = 4). * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: HCMEC (human coronary microvascular endothelial cells) and HCAEC (human coronary artery endothelial cells) were cultured in endothelial cell basal medium MV‐2 (PromoCell, C‐22022) supplemented with the supplement mix (PromoCell, C‐39226).

Techniques: Activity Assay, Incubation, Luciferase

Journal: bioRxiv

Article Title: Mitochondrial COX4I2 drives pericyte-dependent inflammation and emphysema

doi: 10.64898/2026.02.09.703513

Figure Lengend Snippet: Cox4i2 is predominantly expressed in pericytes, smooth muscle cells and fibroblasts a) Schematic illustration of sample acquisition and data generation from CD45 + and CD45 - cells for single-cell RNAseq analyses. b) UMAP representation of CD45 - cells ( n =2–3 per group). c) UMAP highlighting Cox4i2 expressing cells in the pericyte/ smooth muscle cells (SMC) cluster of CD45 - cells. d) DotPlot visualization of marker genes for pericytes, SMCs and fibroblasts (FBs). e) Protein expression of COX4I2 in primary lung FBs, pulmonary artery SMCs (PASMCs), alveolar type II cells (ATIIs), endothelial cells (ECs), pericyte pellet, T cells, neutrophils and macrophages. PASMCs isolated from Cox4i2 −/− mice served as a negative control. f) Track plot of pericyte markers and fibroblast markers g) UMAP representation of the expression of Cox4i2 , Acta2 , or both in pericytes. Red: cells expressing Acta2 ; green: cells expressing Cox4i2 ; yellow: cells expressing both Acta2 and Cox4i2 . h) DotPlot visualization of marker genes expressed in pericytes. i) Labelling of Cox4i2 -expressing cells was performed using Cox4i2 -CreERT2-tdTomato mice. A Kozak-CreERT2-P2A cassette was inserted just before the Cox4i2 start codon, and these mice were crossed with tdTomato reporter mice containing a loxP-STOP-loxP cassette. After tamoxifen (i.p.; TAM) injection, Cre activation removed the stop cassette, enabling tdTomato expression driven by the Cox4i2 promoter. j, k) Representative images. j: tdTomato-positive cells in alveolar structures costained with CD31 as an endothelial marker and ACTA2 as an SMC marker. k: tdTomato-positive cells in pulmonary vessels. Colour code: Red – tdTomato, green – ACTA2, bright cyan – CD31, blue – Hoechst.

Article Snippet: Primary endothelial cells (3□×□10□/mL) in MV-2 basal medium (Promocell) with 0.4% heparin, 0.1% ascorbic acid, and 0.1% hydrocortisone were seeded at 15,000 cells/well.

Techniques: Single Cell, RNA sequencing, Expressing, Marker, Isolation, Negative Control, Injection, Activation Assay