Journal: Cells

Article Title: Differential Dose- and Tissue-Dependent Effects of foxo on Aging, Metabolic and Proteostatic Pathways.

doi: 10.3390/cells10123577

Figure Lengend Snippet: Figure 1. Ubiquitous foxo OE accelerates aging phenotypes in a dose-dependent manner. (a) Survival curves of foxoH and foxoL overexpressing flies and (b) median lifespan of foxo overexpressing flies vs. respective controls [i.e., +/+ (w1118 flies), +/TubGal4 RU486(−) and +/TubGal4 RU486(+) flies]. Statistics of longevity assays are shown in Table S1. (c) Locomotion (climbing activity) of young (Y), middle-aged (M), and aged (A) foxo overexpressing flies vs. controls. (d) Representative CLSM images of intestinal longitudinal muscle fibers’ actin filaments (Phalloidin stain) of female middle- aged foxo overexpressing flies vs. control. Bars, ±SD; n ≥3, ≥10 flies were analyzed per experimental repeat. Statistical significance was measured with unpaired t-test, * p < 0.05, ** p < 0.01.

Article Snippet: Samples were then mounted and viewed using a Digital Eclipse Nikon C1 confocal laser scanning microscope (CLSM) (Nikon Corporation, Tokyo, Japan).

Techniques: Activity Assay, Staining, Control

Journal: Cells

Article Title: Differential Dose- and Tissue-Dependent Effects of foxo on Aging, Metabolic and Proteostatic Pathways.

doi: 10.3390/cells10123577

Figure Lengend Snippet: Figure 2. Perturbation of major bioenergetic and metabolic signaling pathways after ubiquitous foxo OE in Drosophila flies. (a) Representative CLMS images of female adult middle-aged foxo overexpressing, or control flies’ muscle fibers stained with anti-ATP5a (mitochondrial subunit). (b) Relative expression levels of mitochondrial (TFAM, srl/PGC1α, Marf, Drp1, blw/ATP5a) genes in shown transgenic flies. (c) Relative expression levels of Ilp2, Ilp6, ImpL2, InR, Akt1, G6P, GlyP, and GlyS genes (bars are as in b) and, (d) representative immunoblotting analysis showing expression levels of insulin receptor (InR), pAkt1Ser505 (phosphorylated activated form), total Akt1, psggSer21/9 (phosphorylated inhibitory form) and total sgg in indicated foxo overexpressing flies; Gapdh probing was used as a loading reference. (e) Content (%) of TRE (trehalose), GLU (glucose), and GLY (glycogen) in dissected somatic tissues or isolated hemolymph from middle-aged foxo overexpressing transgenic flies (bars are as in b); controls were set to 100%. (f) Representative CLSM images of lipid droplets (LDs) (Bodipy

Article Snippet: Samples were then mounted and viewed using a Digital Eclipse Nikon C1 confocal laser scanning microscope (CLSM) (Nikon Corporation, Tokyo, Japan).

Techniques: Protein-Protein interactions, Control, Staining, Expressing, Transgenic Assay, Western Blot, Isolation

Journal: Cells

Article Title: Differential Dose- and Tissue-Dependent Effects of foxo on Aging, Metabolic and Proteostatic Pathways.

doi: 10.3390/cells10123577

Figure Lengend Snippet: Figure 4. Muscle-targeted foxo OE delays aging dose-dependently. (a) Representative CLSM images of third instar larvae body wall’s muscle fibers after targeted foxo OE in muscles (MhcGal4); samples were stained with an anti-FOXO antibody and counterstained with DAPI. (b–d) Longevity (b), locomotion performance (climbing activity) of young (Y), middle-aged (M), and aged (A) flies (c), and offspring number per laid eggs (d), of Drosophila flies with the shown genotypes vs. respective controls. (e) Representative CLSM images of third instar larvae body wall’s muscle fibers after muscle targeted (Mef2Gal4; driver with strong expression) OE of foxoL; samples were stained with an anti-FOXO antibody and counterstained with DAPI. (f–h) Lifespan (f), locomotion performance (climbing activity) at indicated ages (g), and offspring number per laid eggs (h) of the shown transgenic flies. Statistics of longevity assays are reported in Table S1. Bars, ± SD; n ≥3, ≥10 flies were analyzed per experimental repeat. In (c,d,g,h) significant differences were calculated with the unpaired t-test, * p < 0.05, ** p < 0.01.

Article Snippet: Samples were then mounted and viewed using a Digital Eclipse Nikon C1 confocal laser scanning microscope (CLSM) (Nikon Corporation, Tokyo, Japan).

Techniques: Muscles, Staining, Activity Assay, Expressing, Transgenic Assay

Journal: Cells

Article Title: Differential Dose- and Tissue-Dependent Effects of foxo on Aging, Metabolic and Proteostatic Pathways.

doi: 10.3390/cells10123577

Figure Lengend Snippet: Figure 5. Cardiomyocytes-targeted foxo OE induces heart-tissue remodeling and impacts flies’ physiology. (a) Represen- tative CLSM images of intact young Drosophila heart tubes and of cardiomyocytes following foxoH or foxoL heart-specific (tinC.∆4Gal4) OE; samples were stained with an anti-FOXO antibody and counterstained with DAPI to visualize nuclei. (b) Locomotion (climbing activity) of young (Y), middle-aged (M), and aged (A) flies of the indicated genotypes and (c) longevity of shown transgenic flies after heart-targeted foxo OE vs. controls. Statistics of longevity assays are reported in Table S1. Bars, ± SD; n ≥3, ≥10 flies were analyzed per experimental repeat. In (b) statistical significance was measured with the unpaired t-test, * p < 0.05, ** p < 0.01.

Article Snippet: Samples were then mounted and viewed using a Digital Eclipse Nikon C1 confocal laser scanning microscope (CLSM) (Nikon Corporation, Tokyo, Japan).

Techniques: Staining, Activity Assay, Transgenic Assay