Figure 6.

Double IF of GLB1 and neuronal marker for the hippocampus and the effects of fisetin. (A) Double IF for GLB1 and NEUN to detect senescent neurons. GLB1⁺ cells are stained in green in the cytoplasm (green channel). Very few GLB1⁺NEUN⁺ cells are found in the neurons of the CA 1-4 area. Many GLB1⁺NEUN⁺ cells can be observed in the non-CA area neurons, shown in a distinct patched cytoplasm yellow color (highlighted in the insets). (B) Quantification of GLB1⁺NEUN⁺ cells/total NEUN⁺ cells percentage in the CA1-4 area. (C) Quantification of GLB1⁺NEUN⁺/total NEUN⁺ cells in the non-CA area (large neurons). (D) Double IF for GLB1 and GFAP. GFAP⁺ astrocytes are stained in red, showing several projections in the non-CA area. GLB1⁺ cells are stained green in the cytoplasm. GLB1⁺GAFP⁺ cells are highlighted in the insets. (E) Quantification of GLB1⁺GFAP⁺/total GFAP⁺ cells percentage. (F) Double IF staining of GLB1 and IBA1 to detect senescent microglia. IBA1⁺ cells are stained in red, with smaller size and few projections in the non-CA area. GLB1⁺IBA1⁺ cells are highlighted in the insets. (G) Quantification of GLB1⁺IBA1⁺/total IBA1⁺ cells percentage in non-CA area. Scale bars = 100 µm. White boxes highlight double positive cells in each staining. Exact p values are indicated between the group bars.