Fig. 2

NEK7 kinase activity is required for dendrite and spine morphogenesis. a Fourteen DIV neurons, co-transfected at 7 DIV with control or Nek7 shRNA and with plasmid expressing FLAG-tagged GFP, NEK7, or NEK7 D179A kinase-dead mutant were fixed and stained with anti-FLAG antibody. Scale bar, 50 μm. b Quantification of the total dendrite length in 14 DIV neurons in the conditions shown in (a); n = 3 independent experiments. Total number of neurons: 42 (Control), 33 (Nek7 depleted), 28 (rescue with WT Nek7), and 30 (rescue with Nek7 D179A). *P < 0.05, **P < 0.01, ***P < 0.001, by two-tailed t-test. c Sholl analysis of 14 DIV neurons in the conditions shown in (a). Mean number of intersections are plotted. Statistics as in (b). d Representative confocal images of dendritic spines in primary dendrites of anti-FLAG-stained neurons as in (a). Scale bar, 1 μm. e Quantification of the density of spines as in (d); n = 3 independent experiments, total of 20 to 25 neurons per condition, total number of spines analyzed per condition 366 < n < 893. n.s. non-significant, *P < 0.05, **P < 0.01 by two-tailed t-test. f Quantification of head diameter of spines as in (d). Statistics as in (e). Error bars: s.e.m.