thioflavin S staining was performed to analyse tau tangles in the brains of tauopathy model mice upon MARK4 protein ablation. Briefly, after washing, slices were mounted on a glass slide and allowed to completely dry on a heat plate at 42°C, followed by incubation with 0.5 mM thioflavin S (Merck, T1892) in 50% ethanol for 7 min.35 (link),36 (link) The number of thioflavin S-positive puncta was quantified using default thresholding segmentation and measure-particle functions in ImageJ (U. S. National Institute of Health, Bethesda, Maryland, USA).
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