Fig. 6

zfmlc1 and hMLC1 overexpressed in primary Glialcam^−/− astrocytes are located at cell-cell junctions. a, b Primary astrocytes isolated from Glialcam^−/− mice were incubated for 18 h at 28 °C. Then, MLC1 was detected by immunofluoresce (a) and protein levels were monitored by Western blot (b). Actin served as a loading control. Lack of signal using GlialCAM antibodies confirmed the lack of protein expression. c, d Overexpression using adenoviruses of mlc1 from zebrafish (zfmlc1, c) and human HA-tagged MLC1 (hMLC1, d) detected both MLC1 proteins at cell-cell junctions (arrowheads) in primary astrocytes isolated form Glialcam^−/− mice. We used antibodies detecting the zebrafish MLC1 or the HA epitope, which did not detect the endogenous Mlc1. Scale bar: 10 μm