Research Paper Volume 12, Issue 22 pp 23233—23250

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).