Fig. 4

HDAC6 controls dendrite development and Golgi polarization through cortactin deacetylation. Neurons expressing cortactin rescued dendrite growth (a-e) and Golgi polarization (f, g) in HDAC6 knockdown neurons. a HDAC6 knockdown neurons had abnormal dendrite growth. Neurons expressing HDAC6 shRNA together with cortactin rescued phenotype. Scale bar, 50 μm. b Sholl graphs of neurons expressing Venus or HDAC6 shRNA or HDAC6 shRNA together with cortactin. Data represents average of three independent experiments (n = 10 for each group in each experiment). c-e Graphs represent the number of primary dendrites (c), longest dendrite length (d) and total dendrite length (e). f The Golgi apparatus was stained by GM130 antibody in neurons. Hippocampal neurons expressing HDAC6 shRNA did not polarize Golgi apparatus. But neurons expressing HDAC6 shRNA with cortactin WT restored Golgi polarization. Scale bar, 50 μm. Acetylation defective or mimetic form of cortactin expression changed dendrite development and Golgi polarization. g Quantification of percentage of neurons with dendritic Golgi. Data represents average of three independent experiments (n = 80–110 for each group in each experiment). h-j Acetylation defective or mimetic form of cortactin expression changed dendrite development and Golgi polarization. h Expression of deacetylation mimetic form of cortactin rescued HDAC6 knockdown phenotype. Overexpression of acetylation mimetic form of cortactin influenced dendrite development and Golgi deployment in primary hippocampal neurons. The Golgi apparatus was stained by GM130 (red). Scale bar, 50 μm. i, j Quantification of the number of primary dendrites (i) and the percentage of neurons with dendritic Golgi (j). Data represents average of three independent experiments (n = 40–120 for each group in each experiment). *p < 0.05, **p < 0.01, ***p < 0.001 by two-tailed Student’s t-test. The symbol # represents the number. Error bar means ± S.E.M. in all graphs