Fig. 2

Changes of hippocampal cell numbers in sham and HI mice. A Representative gross morphology of mice Brains at P8, P14, and P60 (Left), and the morphological structure of hippocampus in HI mice at P8, P14, and P60 (Right). Blue: DAPI. A noticeable atrophy of the hippocampus on the ligated side in HI mice can be observed at P60. B Neun immunofluorescence staining of hippocampal tissue sections, showing a significant reduction of Neun-positive neurons in HI mice. Red: Neun, Blue: DAPI, scale bar = 100 μm. C Mean and standard error of the number of Neun-positive neurons (two-tailed unpaired Student’s t-test, n = 5 mice/group). D Iba1 immunofluorescence staining, demonstrating a marked elevation in Iba1-positive microglia in HI Mice. Green: Iba1, Blue: DAPI, white arrows: activated microglial cells. E The number of Iba1-positive microglia (two-tailed unpaired Student’s t-test, n = 5 mice/group). F GFAP immunofluorescence staining, indicating a pronounced increase in GFAP-positive astrocytes in HI Mice. Red: GFAP, Blue: DAPI, white arrows: activated astrocytes. G The number of GFAP-positive astrocytes (two-tailed unpaired Student’s t-test, n = 5 mice/group). Cell counting was limited to the hippocampal region, excluding areas such as the cortex. This applies to all quantifications mentioned in panels C, E, and G