Fig 2: Confocal microscopy of HeLa cells after transfection with DNA2aD1–DNA3rD5 (row 1), DNA2rD1–DNA3aD8 (row 2), DNA2aD2–DNA3aD8 (row 3) and DNA2rD4–DNA3aD8 (row 4). The visualization was performed using a Leica TCS-SPE (DMi8) inverted microscope with an ACS APO 63×/1.30 oil objective. For DNA2aD1–DNA3rD5 ?exc = 405 nm (UV laser), ?em = 435–470 nm (blue) and 575–750 nm (yellow), for DNA2rD1–DNA3aD8 ?exc = 405 nm (UV laser), ?em = 415–550 nm (blue) and 575–750 nm (red), for DNA2aD2–DNA3aD8 ?exc = 488 nm (argon ion laser), ?em = 490–550 nm (green) and 550–675 nm (red), for DNA2rD4–DNA3aD8 ?exc = 488 nm (argon ion laser), ?em = 490–550 nm (green) and 675–800 nm (red), scale bar = 20 µm.

Fig 3: Enhanced expression of GLUT-1 under hypoxia microenvironment. A. Cells were cultured under normoxia (N) and another set in hypoxia (H) for 72 h and stained for GLUT-1 by immunocytochemistry. The Images were taken with DMi8 Leica microscope and quantified using ImageJ. B. Western blotting was done by the SDS-PAGE method and densitometry done using ImageJ. The relative expression was normalized to tubulin. Data was analysed using student t test by graph pad prism. Each value represents the mean ± SD of three independent experiments, * indicates p < 0.05. Scale bar = 50 µm. Representative Western blots are shown as cropped images. Full-length blots are presented in Additional File 3 (Fig. S3)

Fig 4: Senile plaques labeled with Thioflavin-S.1% (green) in the hippocampus of (A–C) senescence-accelerated mice resistant (SAMR)-1, (D–F) Senescence Accelerated Mouse-Prone 8 (SAMP-8), (G–I) Li, (J–L) EE, and (M–O) Li + EE. Cellular nuclei were labeled with DAPI (blue). Images were obtained using an inverted DMi8 microscope (Leica, Wetzlar, Germany) with a 20× objective. Scale bar: 100 µm.

Fig 5: Labeling of NeuN (green) in the CA1 hippocampal region of (A–C) senescence-accelerated mice resistant (SAMR)-1, (D–F) Senescence Accelerated Mouse-Prone 8 (SAMP-8), (G–I) Li, (J–L) EE, and (M–O) Li + EE mice. Cellular nuclei were labeled with DAPI (blue). Images were obtained using an inverted DMi8 microscope (Leica, Wetzlar, Germany) with a 10× objective. Scale bar: 100 µm.