Journal: Aging Cell

Article Title: Ghrelin Alleviates Aging‐Related Cognitive Impairment by Regulating Autophagy‐Related Signaling via the SHBG / JNK1 /Beclin1 Axis in Microglia

doi: 10.1111/acel.70732

Figure Lengend Snippet: The information on the antibody.

Article Snippet: p21 , 1:2000 , 10355‐1‐AP , Rabbit , Proteintech , USA.

Techniques:

Journal: Aging Cell

Article Title: Single‐Cell Mapping Identifies Aging‐Associated Neutrophil Remodeling Across Multiple Organs

doi: 10.1111/acel.70735

Figure Lengend Snippet: Neutrophils from aged mice display enhanced inflammatory and effector‐associated programs. (A) t‐SNE plot of the integrated neutrophil atlas from multiple organs, including blood, intestine, liver, lung, vascular tissue, heart, kidney, and brain, colored by age group. The number of neutrophils contributed by each group is shown in the accompanying bar plot. (B) Volcano plot showing differential gene expression in neutrophils from aged versus young mice. Genes with adjusted p value < 0.05 and log2 Fold Change > 0.5 were designated as differentially expressed genes. (C, D) Hallmark functional enrichment analysis of gene sets enriched in neutrophils from aged (C) or young (D) samples. Dot size represents gene count and color indicates adjusted significance. (E) Sample‐level comparison of marrow proximity and reactive oxygen species (ROS) module scores in neutrophils from young and aged mice. Each dot represents one biological sample. (F) Representative Cytox Green staining images showing NET formation in neutrophils isolated from young and aged mice. Quantification of NET‐positive cells is shown on the right, each data point represents one biological replicate. Scale bar, 50 μm. (G) Representative senescence‐associated β‐galactosidase (β‐Gal) staining images of MOVAS and AML‐12 cells following treatment with conditional medium derived from young or aged neutrophils. Quantification of β‐Gal‐positive cells is shown on the right, each data point represents one biological replicate. Scale bar, 20 μm. (H, I) Immunoblot (H) and its quantification analysis (I) of MMP12, MMP9, P16, and P21 expression in MOVAS and AML‐12 cells treated with conditioned medium from young or aged neutrophils. GAPDH served as a loading control. Relative protein intensities normalized to young group are indicated below each blot. Each plot represents an independent biological replicate. (J) Relative RNA expression of senescence‐associated secretory phenotype (SASP) genes ( Il6 , Tnf , and Ifn ) and extracellular matrix remodeling genes ( Mmp3 and Mmp9 ) in MOVAS cells following treatment with conditioned medium from young or aged neutrophils. Data are presented as mean ± s.e.m. Statistical significance was determined using two‐sided unpaired t ‐test (E–G, I) and two‐way ANOVA followed by Tukey's multiple‐comparisons test (J). * p < 0.05 ; ** p < 0.01; *** p < 0.001.

Article Snippet: MMP2, MMP9, MMP12, P16, and GAPDH were purchased from Proteintech; P53 was purchased from Affinity; P21 was from Santa Cruz.

Techniques: Tissue, Gene Expression, Functional Assay, Analysis, Comparison, Staining, Isolation, Derivative Assay, Western Blot, Expressing, Control, RNA Expression