FIG. 4.

α1ACT transcription factor expression in SCA6 neurons. (A) Nuclear α1ACT is detected in the nuclei of SCA6 and control 5w neurons by immunostaining with antibody 2#. The same antibody was also used in (B–D). Scale bar 50 μm. (B) Quantification of the MAP2-positive neurons expressing α1ACT detected with antibody #2. Each bar represents the mean ± s.e.m. from three independent experiments. One-way ANOVA test performed; no differences were found between genotypes. (C) Representative confocal Z-stack projection images of neurons immunostained with antibody 2#. α1ACT transcription factor immunoreactivity is detected in nuclei. RedDot2 was used for nuclear staining. Scale bar 20 μm. (D) Quantification of nuclear α1ACT immunofluorescence. Graph bar of relative α1ACT immunofluorescence intensity in nuclei. Each bar represents mean ± s.e.m. from three independent experiments. One-way ANOVA test was performed and did not indicate differences between genotypes. (E) The nuclear localization of α1ACT in neurons is confirmed by immunostaining with antibody 3#, designed to detect all known proteins encoded by the CACNA1A gene (Fig. 1A, see Materials and Methods section). Scale bar 20 μm.