Fig. 9.

Effects of NL1 overexpression on BDNF fluorescence intensity and distribution. A, B Overexpressing NL1 increases BDNF immunofluorescence at AZs. Representative images and quantification of BDNF fluorescence intensity in the dendritic region of neurons expressing mOrange or NL1–IRES–mOrange. On DIV5 the neurons were fixed and immunostained for BDNF (green). The average intensity of fluorescent puncta was quantified. Mean ± SEM; N = 3 experiments, n = 10 cells per condition and experiment; Student’s t test: ***p< 0.0001. Scale bar is 10 μm. C, D BDNF immunosignals colocalize with bassoon in NL1 overexpressing neurons. Mean ± SEM; N = 3 experiments, n = 10 cells per condition and experiment; Student’s t test: ***p< 0.0001. Scale bar is 10 μm. E, F STED nanoscopy images showing the distribution of BDNF. E DIV5 cultured hippocampal neurons transfected with NL1-GFP or GFP and immunostained for GFP, bassoon, and BDNF (see labels). F Average number of dendritic BDNF puncta per 1 μm. Mean + SEM; N = 3 experiments, n = 10 cells per experiment and condition; Student’s t test: ***p< 0.0001. Scale bar is 1 μm