Characterization of Glial Cell Models and In Vitro Manipulation of the Neuregulin1/ErbB System
Figure 7
The expression of NRG1 intracellular domain stimulates cell migration. (a) To express the NRG1 intracellular domain (ICD), two constructs were prepared: one containing the nuclear localization sequence (NLS) and one lacking it (ΔNLS). ICD is shown in blue, NLS in yellow, and the inserted ATG in red. (b) Validation of nuclear and cytoplasmic localization of NRG1-ICD fragments. Nuclear and cytoplasmic proteins were extracted from mock (CTR) and NRG1 transfected COS7 cells and subjected to SDS-PAGE and Western blot analysis. Membranes were incubated with anti-NRG1 (sc-348) antibody. Asterisk indicates an unspecific band. (c) NOBEC transiently expressing the NRG1 intracellular domain (ICD), containing (NLS) or lacking (ΔNLS) the nuclear localization sequence, were assayed for migration activity; data show that the cytoplasmic protein confers a migratory activity higher than the migratory activity conferred by the nuclear protein. (d) RT4-D6P2T cells transiently expressing the NRG1 intracellular domain (ICD), containing (NLS) or lacking (ΔNLS) the nuclear localization sequence, were assayed for migration activity. No statistical difference between cells transfected with the empty vector and cells transfected with the two constructs was observed. Values represent the average of three biological replicates performed as technical triplicates. Values of each replicate are expressed in percentage with respect to the total number of cells that migrated in that experiment (*, ; **, ; ***, ).
