Journal: Nature Communications
Article Title: A drug-free cardiovascular stent functionalized with tailored collagen supports in-situ healing of vascular tissues
doi: 10.1038/s41467-024-44902-2
Figure Lengend Snippet: a Representative rhodamine-conjugated phalloidin (TRITC-Phalloidin, red) and 4′,6-diamidino-2-phenylindole (DAPI, blue) fluorescence images of HUVECs showing the (rhCol III/PDA-PEI) n coatings loaded with different amounts of rhCol III all encouraged HUVECs compared with control PLA group. Scale bars, 200 μm. Quantification of ( b , c ) cell number and ( d , e ) cell viability of HUVECs cultured on uncoated and (rhCol III/PDA-PEI) n -coated PLA ( n = 1, 2, and 4) sheets after 1, 3, 5, and 7 days of culture ( n = 5 independent samples). f Volcano plot showing differentially expressed genes in the (rhCol III/PDA-PEI) 2 group compared to the PLA control group. Downregulated and upregulated genes are colored blue and red, respectively, at significantly differentially expressed thresholds │log 2 FC│ > 1.2 and p < 0.05. g Gene Ontology (GO) analysis of differentially expressed genes in PLA versus (rhCol III/PDA-PEI) 2 . h Heatmap of the differentially expressed genes in the PLA and (rhCol III/PDA-PEI) 2 groups. i Circular visualization of the results of gene-annotation enrichment analysis. j Quantification of the expression of representative genes in PLA versus (rhCol III/PDA-PEI) 2 , validated by qRT-PCR arrays. The value in the PLA group was normalized to that in the (rhCol III/PDA-PEI) 2 group ( n = 3 independent samples). k Representative immunofluorescence staining of CCL5 (green), GATA3 (green), XBP1 (green), and CEACAM6 (green) of HUVECs on 3 days of culture. Scale bar, 50 µm. l Schematic diagram of the potential three signaling pathways involved in the regulation of HUVEC behavior induced by the (rhCol III/PDA-PEI) 2 coating, including PI3K/AKT, mTOR, and MAPK. Two-way ANOVA with Tukey’s multiple comparisons was used for the comparisons in ( b )–( e ) and ( j ). Two-sided Student’s t test with multiple testing corrections was used in ( f ) and ( g ). The data are presented as the mean ± SD ( p values < 0.05 were considered statistically significant).
Article Snippet: The primary antibodies used in this study included mouse monoclonal XBP1 (Cat. No.: sc-8015, Clone: F-4, Santa Cruz Biotechnology, USA, 1:50), mouse polyclonal CCL5 (Cat. No.: sc-365826, Clone: A-4, Santa Cruz Biotechnology, USA, 1:50), mouse monoclonal CEACAM6 (Cat. No.: sc-59899, Clone: 9A6, Santa Cruz Biotechnology, USA, 1:50), rabbit monoclonal GATA3 (Cat. No.: ab199428, Clone: EPR16651, Abcam, USA, 1:500), rabbit polyclonal F4/80 (Cat. No.: 29414-1-AP, Proteintech, China, 1:100), rabbit monoclonal CD68 (Cat. No.: ab283654, Clone: EPR23917-164, Abcam, USA, 1:100), rabbit polyclonal CD86 (Cat. No.: bs-1035R, Biosynthesis Biotechnology co., ltd, USA, 1:200), rabbit monoclonal CD206 (Cat. No.: 24595, Clone: E6T5J, Cell Signaling Technology, USA, 1:200), mouse monoclonal α-SMA (Cat. No.: ab7817, Clone: 1A4, Abcam, USA, 1:200), and rabbit monoclonal MMP2 (Cat. No.: 10373-2-AP, Clone: SB13a, Proteintech, USA, 1:200), mouse monoclonal CD31 (Cat. No.: ab9498, Clone: JC/70A, Abcam, USA, 1:200), and rabbit polyclonal eNOS (Cat. No.: ab5589, Abcam, USA, 1:100).
Techniques: Fluorescence, Control, Cell Culture, Expressing, Quantitative RT-PCR, Immunofluorescence, Staining, Protein-Protein interactions