Skip to main content
Figure 4 | Molecular Neurodegeneration

Figure 4

From: Histone deacetylase-3 interacts with ataxin-7 and is altered in a spinocerebellar ataxia type 7 mouse model

Figure 4

Role of HDAC3 deacetylase activity in ataxin-7 post-translational modification and subcellular localization. (A) Ataxin-7-10Q, ataxin-7-10Q (1–266), ataxin-7-92Q or ataxin-7-92Q (1–266) were cotransfected with vector control or HDAC3 or a catalytically-inactive mutant of HDAC3, HDAC3 (H134A/H135A), in HEK293T cells. Western blot analysis of cellular lysates with ataxin-7 (PA1-749) antibody revealed the presence of bands corresponding to the wild type, expanded and stop mutant ataxin-7. Bands of higher molecular weight that are detected when ataxin-7 is co-expressed with HDAC3 (indicated by asterisks) are also present when ataxin-7 is co-expressed with the catalytically-inactive HDAC3 mutant. Co-expression of HDAC3 was confirmed by immunoblotting (HDAC3 antibody sc17795; data not shown) and GAPDH was used as a loading control. (B) Immunocytochemistry of HEK293T cells transiently transfected with ataxin-7-10Q (wild type) or ataxin-7-92Q (expanded), and co-transfected with HDAC3 or the catalytically-inactive mutant HDAC3 (H134A/H135A). Cells were fixed and stained with antibodies to ataxin-7 conjugated to Alexa 488 (green), HDAC3 conjugated to Alexa 555 (red), and counterstained with DAPI (blue). Cells were imaged using confocal microscopy to elucidate colocalization of ataxin-7 and HDAC3, which can be detected by yellow co-staining. Scale bar represents 5 μm wild type.

Back to article page