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. Author manuscript; available in PMC: 2020 Jun 29.
Published in final edited form as: Cell Rep. 2020 May 26;31(8):107685. doi: 10.1016/j.celrep.2020.107685

Figure 2. TGF-β-Enhanced Interaction of Ankyrin-G and Usp9X Regulates Spine Morphogenesis.

Figure 2.

(A) SIM image of mCherry-expressing neurons (red) with treatment of vehicle or TGF-β (20 ng/mL) for 1 h to detect the interaction between ankyrin-G and Usp9X by PLA (cyan). Scale bar, 10 μm, top. Magnified image of boxed spine and dendrite (bottom). Scale bar, 2 μm, bottom.

(B) The number of PLA puncta per 10 μm in neurons treated with vehicle or TGF-β. Bars also show the distribution of PLA puncta in spines (white) and dendrites (gray) (vehicle, n = 28 neurons; TGF-β, n = 33 neurons). **p < 0.01; ***p < 0.001 by two-tailed unpaired t test.

(C) Confocal images of neurons transfected with control RNAi (control) or shRNA against Usp9X (shUsp9X) or ankyrin-G (shAnkG) and subsequently treated with vehicle (top) or TGF-β (20 ng/mL) (bottom) for 1 h. Scale bar, 5 μm.

(D and E) Spine head area (D) and density (E) in control (black bars), shUsp9X (red bars), and shAnkG (blue bars) with vehicle (plain pattern) or TGF-β (comb pattern) (n = 20 neurons for each group). *p < 0.05; ***p < 0.001; †††p < 0.001 by one-way ANOVA followed by non-parametric statistical analysis. All data are presented as mean ± SEM.