Fig. 5

Validation of metallothionein, mitochondrial changes and SNORD114-6 with IHC or in situ. A Representative images of metallothionein levels the control, AD and resilient groups. B Quantification of metallothionein immunoreactivity (MT-ir). MT-ir is significantly increased in resilient compared to control cases (A–B: F(2,22) = 5.29, p = 0.0008, AD versus control; p = 0.964, resilient versus control; p = 0.001, resilient versus AD; p = 0.003) C Representative images of mitochondria using MT-CO1. D Quantification of the proportion of OD from MT-CO1 in and outside of the soma. The proportion of MT-CO1 outside compared to inside of the soma is significantly larger in the resilient compared to the control cases (F(2,22) = 3.87 p = 0.036, AD versus control; p = 0.387, resilient versus control; p = 0.028, resilient versus AD; p = 0.299). E Fluorescent IHC stains show that metallothionein (green) is present in GFAP-positive astrocytes (red). Double-labeled astrocytes are denoted with an arrow, nuclei are stained with DAPI (blue). F–G MTCO1 (green) is primarily present in NeuN-positive neurons (red) and less in Iba1-positive microglia (white) or GFAP-positive astrocytes (red). Double-labeled astrocytes are denoted with an arrow and neurons with an asterisk, nuclei are stained with DAPI (blue). H Normalized counts of SNORD114-6 in the different groups. I In situ signal of SNORD114-6 in the nucleolus in neurons. J Relative gene expression of SNORD114-6 with qPCR. There are no statistical differences between the groups. Scale bars in all panels are 50 µm. Data of IHC is represented as average ± SEM, in situ as relative gene expression to housekeeping genes. p < 0.05: *, p < 0.01: **