Figure 8

Gβγ interacts with MTs in primary hippocampal and cerebellar neurons. Neuronal primary cultures from hippocampus (A, B) and cerebellum (C, D) of rat brains were prepared as described in the methods. Hippocampal (A) and cerebellar (C) neurons were processed for confocal microscopy using anti-tubulin (red) and anti-Gβ (green) antibodies. Areas of overlay appear yellow. The enlarged view of the white boxes (c’, f’) depicts Gβγ-tubulin co-localization in the neuronal process in hippocampal and cerebellar neurons. The scale bar is 20 μm. Microtubules (MT) and soluble tubulin (ST) fractions were prepared from hippocampal (B) and cerebellar (D) neurons as described in the methods. Equal amount of proteins from each fraction were subjected to co-immunoprecipitation using anti-Gβ antibody or in the absence of primary antibody (No ab) followed by an immunoblot analysis of immunoprecipitates (IP) and supernatants (SUP) using anti-α-tubulin antibody (B, D).