Exploratory experiment designed to discover new patterns targeting TFEB in NSC34 neuroblastoma x spinal cord cells. Primary outcome: TFEB nuclear translocation and lysosomal membrane permeabilization
This experiment investigated the mechanism by which trehalose induces autophagy in cellular models of motoneuron degeneration. The study demonstrated that trehalose causes rapid and transient lysosomal enlargement and membrane permeabilization (LMP), which correlates with calcium-dependent phosphatase PPP3/calcineurin activation, leading to TFEB dephosphorylation and nuclear translocation. The researchers used immunofluorescence microscopy to track lysosomal changes and TFEB subcellular localization, and performed biochemical assays to measure phosphatase activity and protein modifications.
Cells treated with trehalose, followed by immunofluorescence analysis for TFEB localization and lysosomal markers, biochemical assays for PPP3/calcineurin activity, and measurement of lysosomal membrane integrity
Trehalose treatment should induce lysosomal changes leading to TFEB activation and autophagy upregulation
Significant TFEB nuclear translocation, lysosomal enlargement, and increased autophagy gene expression
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