Figure 5. Metformin alleviates the depolymerization of MTs and neurites retraction by increasing NEAT1. (A, B) Quantitative PCR analysis of NEAT1 and FZD3 expression of 0 to 1mM metformin treatment on SH-SY5Y cells for 48h. (C) The NEAT1 expression was detected in shNEAT1v2 cells and shCtrl cells after treated with 0 to 1mM metformin. (D) The NEAT1v2 siRNA and Ctrl siRNA transfected SH-SY5Y cells were treated with 0.4mM metformin. Immunofluorescence staining were detected with antibody against α-tubulin(red) and subjected to confocal microscopy analysis. DAPI (blue) was used to stain the nuclei. Scale bars 50μm. Image J software was used to count the cell length. (E) Protein levels of FZD3, P-GSK3β, GSK3β, p-tau(ser396) and GAPDH were detected by immunoblotting after 48h treatment of 0.4mM metformin in NEAT1v2 siRNA and Ctrl siRNA transfected SH-SY5Y cells. (mean ± s.d, *P < 0.05, Student 2-tailed t test).