Figure 4
WT Syn increases MT dynamics in neuronal cells.
(a) Representative western blot showing the levels of expression of human Syn (Syn) in 5 days NGF-differentiated control (GFP) and transfected (Syn) PC12 cells. The antibody used (Sigma-Aldrich) recognized both human and rat Syn. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is used as loading reference. (b) Representative micrographs of double transfected PC12 cells differentiated 3 days with NGF, showing the EB3 comets (EB3-mCherry, inverted contrast) and, in the inset, the GFP channel (GFP, grey line marks the cell boundary). The grey box indicates the area from which the frames of the time series were taken (elapsed time is shown). Scale bar, 5 μm. Box plots of the number (c, *BASAL = 0.007, *RECOVERY = 0.005), speed (d, *BASAL = 0.02, *RECOVERY = 0.007) and lifetime (e, *BASAL = 0.02, *RECOVERY = 0.02) of MT growths, in each experimental condition. BASAL indicates cell cultures maintained at 37 °C whereas RECOVERY indicates registration during the rewarming phase after 30 min at 4 °C. At least 30 cells were analysed per condition, for a total number of 2200 tracks, or more. *p < 0.05 vs GFP, according to Student’s t-test. The analyses of the relative neurite’s area are reported in Supplementary Fig. S5.
