Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 3 | The Effect of SAA1 on old-aged tissues. a Schematic drawing of the effect of SAA1 on tissue. b Young fibroblasts or c smooth muscle cells (PASMCs) were treated with rhSAA1 for 24 h and SASP expression was analyzed using real-time PCR (left panel). MMP9 protein expression was analyzed by ELISA (right panel). d Serially dissected human colon tissues were subjected to IHC analysis for SAA1 and MMP9. Corresponding quantification data is shown in the right panel. IHC results were presented as the percentage of positive cells in the stromal region. e PASMCs were treated with rhSAA1 at the indicated concentration for 24 h and real-time PCR was performed using primers for atrophy-related genes. f IHC ana- lysis for SAA1 in colon tissue from young and elderly subjects, and muscular mucosa thickness was measured. g IHC analysis for type IV collagen of colon tissues from young and elderly subjects is shown. “1” and “2” indicate high-magnification views of the original figure (left panel). Each protein expression was presented as weak, moderate, and strong (right upper panel). mRNA expression level of COL4A1 and COL4A2 in fibroblasts/smooth muscle cells was analyzed between young and old subjects in scRNA-seq data set (GSE178341)34 (right lower panel). h COL4A1,

Article Snippet: Mouse Serum Amyloid A (SAA) Quantikine ELISA Kit (MSAA00, R&D systems) was used for quantitative measurement of circulating SAA levels according to the manufacturer’s instructions.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Concentration Assay, Lysis

Journal: Nature communications

Article Title: Mid-old cells are a potential target for anti-aging interventions in the elderly.

doi: 10.1038/s41467-023-43491-w

Figure Lengend Snippet: Fig. 6 | SLIT2 has an impact on mid-old cells. a Mid-old cells were treated for 24 h with rhSLIT2, and inflammatory gene expression was analyzed using real-time PCR (upper panel). IL1β and SAA1 protein level was analyzed by ELISA in rhSLIT2-treated mid-old cells (lower panel). b Mid-old cells were treated with rhSLIT2 for the indi- cated times and analyzed for Pyk2-NFκB signaling by western blot analysis (left panel). The number of nuclear p-NFκB-positive cells was quantified in mid-old cells using IF staining (right panel). c SLIT2-expressing lentivirus infected into mid-old fibroblasts, and inflammation related genes expression including SAA1 and IL1β were analyzed using real-time PCR (right lower panel). IL1β and SAA1 protein level was analyzed by ELISA in SLIT2-overexpressing mid-old cells (left lower panel). d The morphology and cell size of mid-old cells were analyzed after treating rhSLIT2 for 20 days. Cell growth rate was analyzed every 4 days for 20 days by counting the number of cells. e The expression of p53 and p21Waf1 were measured in mid-old cells after administration of rhSLIT2 at the indicated concentration for 2 days using real-time PCR (upper panel) and western blot analysis (lower panel). f The changes in gene expression of p53 and p21Waf1-regulating genes were eval- uated after treating cells with rhSLIT2 using RNA-seq FPKM count (left panel). The mRNA expression of SOX2 and OCT4 were analyzed using real-time PCR (right

Article Snippet: Mouse Serum Amyloid A (SAA) Quantikine ELISA Kit (MSAA00, R&D systems) was used for quantitative measurement of circulating SAA levels according to the manufacturer’s instructions.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Staining, Expressing, Infection, Concentration Assay, RNA Sequencing