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. 2018 Apr 9;293(21):8196–8207. doi: 10.1074/jbc.RA117.001613

Figure 6.

Figure 6.

VEGFD treatment prevents the simplification of dendritic morphology promoted by nuclear HDAC4. A, quantification of the total dendritic length of hippocampal neurons at DIV10. Cells were co-transfected with hrGFP and FLAG-tagged constructs lacZ, HDAC4 WT, and HDAC4 3SA at DIV8. HDAC4 3SA versus HDAC4 WT, p = 3 × 10−4; HDAC4 3SA versus lacZ, p = 1 × 10−3; lacZ versus HDAC4 WT, p = 0.94. B, total number of intersections derived from Sholl analysis at DIV10. HDAC4 3SA versus HDAC4 WT, p = 2 × 10−4; HDAC4 3SA versus lacZ, p = 2.1 × 10−3; lacZ versus HDAC4 WT, p = 0.76. C, representative images of cultured hippocampal neurons transfected as in A and treated or not with rVEGFD (100 ng/ml) for 3 days. FLAG tags were stained with Alexa 594. Scale bar is 40 μm. D, quantification of the total dendritic length of hippocampal neurons transfected as indicated, with or without treatment for 3 days with rVEGFD. HDAC4 3SA versus lacZ, p = 1.9 × 10−3; HDAC4 3SA versus HDAC4 WT, p = 1 × 10−4; HDAC4 3SA versus HDAC4 3SA + rVEGFD, p = 0.03; HDAC4 3SA + rVEGFD versus lacZ + rVEGFD, p = 0.14; HDAC4 3SA + rVEGFD versus HDAC4 WT + rVEGFD, p = 0.61. E, total number of intersections derived from the Sholl analysis shown in F. HDAC4 3SA versus lacZ, p = 2.2 × 10−3; HDAC4 3SA versus HDAC4 WT, p = 1 × 10−4; HDAC4 3SA + rVEGFD versus lacZ, p = 0.69; HDAC4 3SA + rVEGFD versus lacZ + rVEGFD, p = 0.15; HDAC4 3SA + rVEGFD versus HDAC4 WT, p = 0.06; HDAC4 3SA + rVEGFD versus HDAC4 WT + rVEGFD p = 0.57; lacZ versus lacZ + rVEGFD, p = 0.98; HDAC4 WT versus HDAC4 WT + rVEGFD, p = 0.76. F, Sholl analysis of hippocampal neurons transfected as indicated, with or without treatment for 3 days with rVEGFD. In total, 8 (lacZ), 9 (HDAC4 WT), and 12 neurons (lacZ + rVEGFD, HDAC4 WT + rVEGFD, HDAC4 3SA, HDAC4 3SA + rVEGFD) from 3 independent experiments were analyzed for each construct (D, E, and F). At DIV10, 12 (lacZ and HDAC4 3SA) and 13 (HDAC4 WT) neurons from 3 independent experiments were analyzed for each construct (A and B). Statistically significant differences were determined by one-way ANOVA (A, B, D, and E) and two-way ANOVA (F) followed by Tukey's post hoc test. ***, p < 0.001; **, p < 0.01; *, p < 0.05. For scatter plots, each point represents a value derived from one neuron. Graphs represent mean ± S.D.