FIGURE 2:

Overexpression of Fign reduces tyr-MTs in astrocytes. (A) Representative images of severing MT. LV-5-Fign lentivirus was introduced into astrocytes for 3 d and immunostained by anti–α-tubulin, and LV-5-katanin lentivirus was used as a functional control of severing MTs. No MT severing was observed, and no short MT fragments were found in LV5-Fign–treated astrocytes, but severing MTs were found in the group of LV5-katanin-P60–overexpressed cells (left, bottom; bar, 20 μm). Right, bottom, magnification of severed MTs. (B) Top, representative Western blotting result showing that LV5-Fign–overexpression cultures have a lower ratio of tyr-tubulin to α-tubulin than LV5-NC cultures. Bottom, statistical data showing a decrease of tyr-tubulin protein after Fign overexpression. The data are shown as mean ± SE. The relative tyr-tubulin protein level (α-tubulin as the control) of LV-NC–overexpressed astrocytes was normalized as 1. LV5-Fign = 0.48 ± 0.04, n = 3, *p < 0.05. (C) Effect of LV5-Fign overexpression on modified tubulins. Astrocytes treated with LV5-NC (negative control; NC) or LV5-Fign lentivirus for 3 d were fixed and immunostained with tyr-tubulin (red) and α-tubulin (green). No short MTs were detected in Fign-overexpression groups for LV5-Fign (bottom) but decreased significantly in the cell edge with tyr-tubulin (white arrows) compared with the LV5-NC–treated astrocytes (top). Bar, 20 μm.