Fig. 4
From: Decreased nuclear Pten in neural stem cells contributes to deficits in neuronal maturation
Ptenm3m4 NSC differentiation is precocious yet abortive, leading to increased immature neuron population. a Representative immunofluorescence staining defines immature neuron population (Tuj1+ cell in green: DAPI+ nuclei in blue) at 3, 5, 7, and 10 days in vitro in the absence of growth factors. b Quantification of staining in a, where a significant increase in Tuj1+ neurons at 5 DIV (p value = 0.05) and 7 DIV (p value = 0.0013) comparing Ptenm3m4/m3m4 to Ptenwt/wt as assessed by Kruskal-Wallis testing with Dunn’s multiple comparison testing. c Immunofluorescence staining for mature neuron (NeuN+ cells in red: DAPI+ nuclei in blue) population in culture for 5, 7, 10, and 14 DIV in the absence of growth factor. d Quantification of mature neurons in c (NeuN+ cells). Kruskal-Wallis testing with Dunn’s multiple comparison testing found significant decreases in NeuN+ cells at 7 (p value = < 0.0001), 10 (p value = < 0.0001), and 14 DIV (p value = 0.003) between Ptenm3m4/m3m4 and wildtype NSCs and significant decreases in NeuN+ cells at 7 (p value = 0.018) and 14 DIV (p value = 0.029) between Ptenwt/m3m4 and wildtype NSCs. e NeuN+ cells to Tuj1+ cells at 7 and 10 DIV (n = 3). Mann Whitney U testing found a significant different between the 7 and 10 DIV NeuN+/Tuj1+ ratios in Ptenm3m4/m3m4 NSCs (p value = 0.0012). f Tuj1+ neurites showing arbored branched intersections of neurites (n = 3). g Simple neurite tracing quantification by Image J (Sholl analysis), showing increased mean length of longest NSC neurite in Tuj1+ cells derived from Ptenm3m4/m3m4 NSCs (means wt vs het vs mut: 165.4 μm vs 215.5 μm vs 305.7 μm), indicating a significant increase between Ptenm3m4/m3m4 and wildtype NSCs (p value = 0.0006) and between Ptenwt/m3m4 and wildtype NSCs (p value = 0.020). h Quantification of branch intersection of neurites, showing consistently increased branch complexity in Ptenm3m4/m3m4 NSCs at increasing distance from nucleus (*p value < 0.05; **p value < 0.01; ****p value < 0.0001)