Melanoma cell adhesion molecule-positive mesenchymal stromal cells alleviate acute respiratory distress syndrome via nuclear factor kappa-B-mediated paracrine regulation

Figure 5 Transcriptomic analysis of mesenchymal stromal cells and validation. A: Heatmap displaying genes differentially expressed between CD146+ mesenchymal stromal cells (MSCs) and CD146- MSCs; B: Kyoto Encyclopedia of Genes and Genomes enrichment analysis demonstrating that differentially expressed genes are enriched in distinct signaling pathways between CD146+ MSCs and CD146- MSCs; C: Gene set enrichment analysis was performed on the differentially expressed genes; D: Cyclooxygenase 2 (COX2) and nuclear factor kappa B (NF-κB) mRNA levels in CD146+ MSCs and CD146- MSCs were quantified by reverse transcription-quantitative polymerase chain reaction; E: Western blot analysis of COX2, NF-κB, and p-NF-κB protein levels; F: COX2 and NF-κB mRNA levels in CD146+ MSCs treated with caffeic acid phenethyl ester (CAPE) for 24 hours were detected by reverse transcription-quantitative polymerase chain reaction. The band density was compared to that of the control and normalized to that of GAPDH using densitometry; G: The protein levels of COX2, NF-κB, and p-NF-κB were detected by western blot in CD146+ MSCs treated with CAPE for 24 hours. Band density was compared to that of the control and normalized to that of GAPDH via densitometry; H: Enzyme-linked immunosorbent assay analysis of hepatocyte growth factor, prostaglandin E2, angiopoietin-1, and vascular endothelial growth factor expression in CD146+ MSCs after treatment with CAPE for 24 hours. Data are presented as the means ± SD and were analyzed by two-sided Student’s t tests. n = 3. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001. KEGG: Kyoto Encyclopedia of Genes and Genomes; TNF: Tumor necrosis factor; PI3K: Phosphoinositide 3-kinases; IL: Interleukin; TGF: Transforming growth factor; MSC: Mesenchymal stromal cell; NF-κB: Nuclear factor kappa B; COX2: Cyclooxygenase 2; CAPE: Caffeic acid phenethyl ester; HGF: Hepatocyte growth factor; PGE2: Prostaglandin E2; Ang-1: Angiopoietin-1; VEGF: Vascular endothelial growth factor.