Fig. 2
From: Decreased nuclear Pten in neural stem cells contributes to deficits in neuronal maturation
NSCs with Ptenm3m4 mutations show decreased nuclear and global Pten levels. a Representative immunofluorescence staining for Pten at 1 DIV (n = 3). Top panel is low magnification (scale bar 25 μm). Bottom panel is high magnification (scale bar 10 μm). b Volocity software-assisted quantification of Pten immunofluorescence, calculating the volume of Pten protein (μm3) in different NSCs compartments at 1 DIV (n = 3). Volumes were converted to percentages of the whole. c Quantification of Pten immunofluorescence staining, showing percentage of Pten-positive nuclei from Ptenwt/wt (n = 4), Ptenwt/m3m4 (n = 4), and Ptenm3m4/m3m4 (n = 3) NSCs at 0, 3, and 5 DIV in the absence of growth factor. Kruskal-Wallis testing, accompanied by Dunn’s multiple comparison post hoc testing, found a significant difference between wildtype and Ptenm3m4/m3m4 NSCs at 3 (p value = 0.035) and 5 DIV (p value = 0.0051). d Representative Western blot analysis for Pten in nuclear and cytoplasmic fractions derived from NSCs. Hsp90 was used as a cytoplasmic protein marker. e Representative Western blot analysis of total Pten expression in Ptenm3m4 NSCs (n = 3) after 1 DIV. f Representative Western blot analysis of total Pten expression in Ptenm3m4 NSCs at three timepoints over 5 days of random differentiation compared (n = 3). g Representative Western blot analysis of Pten, p110α (Pi3k), p-mTOR, and p-Akt in Ptenm3m4 NSCs (N = 3) (*p value < 0.05; **p value < 0.01; ****p value < 0.0001)