human aortic ecs haecs  (PromoCell)

Journal: Redox Biology

Article Title: Big data analytics for MerTK genomics reveals its double-edged sword functions in human diseases

doi: 10.1016/j.redox.2024.103061

Figure Lengend Snippet: Proteomics in HAECs with MerTK gene knockout or control. ( A ) MerTK expression in HAECs incubated with apoptotic Jurkat cells for 1 h. ( B ) Immunochemical staining for MerTK expression in the aortic arch from WT mice. ( C ) Efferocytosis of apoptotic Jurkat cells by HAECs after 1 h of co-incubation. P < Apoptotic Jurkat cells were labeled with green PKH67 (Sigma) and HAECs were labeled with red PKH26 (Sigma). Green cells are apoptotic Jurkat cells that were not engulfed by HAECs. Green/red small round cells are apoptotic Jurkat cells that were engulfed by HAECs. Large red cells are HAECs. (D) Volcano plot illustration in MerTK KO vs. control. Relative protein abundance (log2) plotted against significance level (-log10 P-value), showing significantly (p < 0.05) downregulated (blue), upregulated (red) or non-differentially expressed proteins (grey). (E) Graphic summarization for pathways in MerTK KO vs. control. (F) MerTK KO activates apoptosis signaling. (G) Canonical pathway analysis in MerTK KO vs. control. Color depends on z-score. Blue signifies negative value; orange signifies positive value; and grey signifies no activity pattern. Size is proportional to the number of genes that overlap the pathway. (H) Machine learning analysis for activated or inhibited disease pathways. (I–K) IPA prediction shows that MerTK KO activates premature aging, kidney failure and heart failure. Proteomics data were analyzed by IPA. Data were analyzed with GraphPad Prism 9.4.1 and shown as the mean ± SD (n = 3–5). P < 0.05 was considered statistically significant. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary human aortic ECs (HAECs) and the human Jurkat cell line were purchased from ATCC (Manassas, VA, USA).

Techniques: Gene Knockout, Control, Expressing, Incubation, Staining, Labeling, Quantitative Proteomics, Activity Assay

Journal: Nature Aging

Article Title: Gut microbiota-dependent increase in phenylacetic acid induces endothelial cell senescence during aging

doi: 10.1038/s43587-025-00864-8

Figure Lengend Snippet: a , Schematic of quantification of plasma PAA and PAGln in young (3-month-old) and aged (>24-month-old) C57BL/6J male and female mice. b , c , Plasma PAA ( b ) and PAGln ( c ) levels were measured by LC–MS/MS in mice ( n = 6). d , Correlation between plasma PAA (left) and PAGln (right) concentrations and chronological age in the TwinsUK cohort ( n = 7,303, male and female). e , Schema for PAA production from dietary phenylalanine (Phe) via the bacterial VOR/PPFOR system. f , Shotgun metagenomics workflow for analyzing mouse fecal microbiomes. g , Distribution of VOR and PPFOR gene homologs (%) in microbiomes of aged and young mice ( n = 5–6). KOO169, VOR; KOO179, PPFOR). h , Bar plot depicting age-associated abundance of fecal microbiota profiles at strain level (dark colors represent taxa harboring VOR or PPFOR homologs). The leftmost bar plot demonstrates the proportion of VOR, PPFOR or VOR + PPFOR detected in taxa enriched in aged ( n = 44) versus young ( n = 37) mice. i , Heatmap shows correlations between plasma PAA or PAGln levels and gut bacteria among the top enriched taxa in aged versus young mice ( n = 5–6). j , Correlation between plasma PAA (left) and PAGln (right) concentrations and abundance (%) of Clostridium taxa in human participants ( n = 900, TwinsUK; male and female). k , PAA (top) and PAGln (below) concentrations in the supernatants of anaerobic cultures from Clostridium sp. ASF356 ( n = 8). l , Schematic of ex vivo force tension myography. Aortic rings exhibit vasorelaxation responses (%) to acetylcholine (Ach) ( n = 10). m , SA-β-gal staining of ECs from the ascending aortas of mice ( n = 5–6) and quantification of SA-β-gal + cells (%). n , o , Immunoblots and immunofluorescence represent the expression of p16 INK4a ( n ) and VCAM1 ( o ) in aortic ECs ( n = 5–6). p , γ-H2A.X immunostaining in CD31 + ECs ( n = 5–6). Scale bars, 20, 50 and 100 μm. Error bars represent s.d. ( b , c , m , n ) or s.e.m. ( g , k , l ) or 95% confidence intervals ( d ). Statistical analysis was performed using a two-tailed unpaired Student’s t -test ( b , c , l – n ), two-tailed Pearson correlation analysis ( d ), two-tailed Mann–Whitney U -test ( g , k ), ANCOM method for microbial abundance analysis ( h ), two-sided Spearman’s rank correlation test ( i ) and linear mixed model CLR transformation ( j ). Data are shown as median with min–max; boxes represent interquartile range (IQR); center lines represent the median; whiskers extend from the min to max values ( g , k ). Images created with BioRender.com ( a , d – f , l ). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Primary human aortic ECs (HAECs; Lonza, CC-2535) were cultured in EBM-2 medium (Lonza, 00190860) supplemented with EGM-2 (10% FBS, 2 mM l -glutamine, 100 μg ml −1 penicillin–streptomycin, Lonza, CC-3162/6) at 37 °C, 5% CO 2 .

Techniques: Clinical Proteomics, Liquid Chromatography with Mass Spectroscopy, Bacteria, Ex Vivo, Staining, Western Blot, Immunofluorescence, Expressing, Immunostaining, Two Tailed Test, MANN-WHITNEY, Transformation Assay