Figure 3

Treatment with CP2 prevents aggregate formation caused by expression of truncated form of mhtt in neurons and glial cells. A. Transfection with truncated N-terminal form of mhtt with 82 polyglutamines (GFP-HD82) causes formation of aggregates (arrows) in striatal neurons (a) and glial cells (b) from control mice. Expression of htt fragment with 19 polyglutamines (GFP-HD19) does not cause formation of aggregates (e, f). Pre-treatment with 2 μM of CP2 prevents formation of aggregates in cells transfected with GFP-HD82 (c, d). Images of live cells were taken using LSM 510 confocal microscope with 100× oil DIC lens (1.4 na). Scale bar, 10 μm. B. Quantification of aggregate formation in neurons and glial cells from control mice with and without CP2 treatment. C. Treatments with TP compounds do not affect the expression of mhtt in primary neurons from HD mice. Neurons (E17) from HD mouse were treated with different doses of CP2, TP3, and TP4 for 6 days. Western Blot analyses were performed using specific monoclonal anti-huntingtin antibody 2166 (1:3000, Chemicon). Lane 1 – HD neurons, no treatment; lane 2 – HD neurons treated with 2 μM CP2; lane 3 – HD neurons treated with 5 μM CP2; lane 4 – HD neurons treated with 2 μM TP3; lane 5 – HD neurons treated with 2 μM TP4. Anti-tubulin antibody (DM1A, 1:6000, Sigma) was used as loading control.