NGL-2 controls the spatial polarization of stabilized microtubules. A, NGL-2 overexpression increased the relative intensity of acetylated α-tubulin in multiple neurites of neurons. Hippocampal neurons were transfected with pEGFP-NGL-2 or pEGFP-empty vector. Transfected neurons were identified by EGFP. Cells were fixed and stained with anti-acetylated α-tubulin antibodies, a stabilized microtubule marker, and anti-α-tubulin. Scale bar, 25 μm. B, Quantification of the relative fluorescence intensity of acetylated α-tubulin vs α-tubulin. The error bars represent the SD. *p < 0.001, Student's t test. N.S., Not significant. C, NGL-2 shRNA decreased the relative fluorescence intensity of acetylated α-tubulin in all neurites of neurons. Hippocampal neurons transfected with NGL-2 shRNA or control shRNA were identified by GFP. Cells were fixed and stained with anti-acetylated α-tubulin antibodies and anti-α-tubulin. Scale bar, 25 μm. D, Quantification of the relative intensity of acetylated α-tubulin vs α-tubulin. The error bars represent the SD. *p < 0.001, Student's t test. N.S., Not significant.