Fig. 4

KLHL17/AF knockdown impairs dendritic spine maintenance and decreases F-actin intensity at dendritic spines. Cultured hippocampal neurons were transfected with the indicated plasmids at 22 DIV and harvested for immunostaining at 28 DIV. a–c AF knockdown impairs dendritic spine maintenance. AF knockdown reduces dendritic protrusion width, but not protrusion length or density. The AF-rescue construct AF(res) rescues the defect of dendritic spine morphology caused by AF-miR. a Representative images. b Quantification of protrusion density. c Quantification of the width and length of dendritic protrusions shown. d–f AF knockdown reduces the dendritic F-actin signal intensity in dendritic spines. Phalloidin staining was performed to label F-actin. d Representative images of whole cells and enlarged dendrite segments. e Representative images of dendritic protrusions. Heat maps show the intensities of F-actin. f Quantification of F-actin intensity at dendritic protrusions. Samples were randomly collected from two independent experiments without knowing the treatment. N the number of analyzed dendrites; n, the number of analyzed dendritic protrusions. Data represent the mean plus SEM. ***P < 0.001; ns not significant. Scale bars: a 5 μm; d whole cell, 20 μm; enlarged inset, 2 μm; e 1 μm. One-way ANOVA (b, f); Kolmogorov–Smirnov test for cumulative probability (c)