Apo tirf objective

All images were taken with a spinning-disk (Yokogawa CSU-X1) confocal microscope with a 100X oil immersion Apo TIRF objective (NA 1.49) and an Andor DU-897 EMCCD camera on a Nikon Eclipse Ti body. A 488nm laser for imaging GFP and global activation, and a 561 was used laser for imaging mCherry. The imaging chamber was maintained at 37°C and 5% CO2 (Okolab) with a 96 well plate adaptor. Local activation was performed by using a Mightex Polygon digital micromirror device (DMD) to pattern blue light (488nm) stimulation from a Lumencor SpectraX light engine. All image acquisition was performed using Nikon Elements Advance Research software.

Images of Corelets, nucleoli, and nuclear speckles were taken with a spinning-disk (Yokogawa CSU-X1) confocal microscope with a 100X oil immersion Apo TIRF objective (NA 1.49) and an Andor DU-897 EMCCD camera on a Nikon Eclipse Ti body. A 488nm laser for imaging GFP and global activation, and a 561 was used for imaging mCherry. The imaging chamber was maintained at 37°C and 5% CO 2 (Okolab) with a 96 well plate adaptor. Z-stacks were taken every 300nm for 15 μm using an ASI MS-2000 stage controller. For drug treatments, DRB (Sigma) or ActD (Sigma) were dissolved in DMSO at 50mg/mL and 2mg/mL stock concentrations respectively, which was diluted in complete DMEM. Cells were treated by replacing media and incubating for 4 hours before imaging.
Images of Htt-PolyQ were taken on a Nikon A1 laser scanning confocal microscope using a 60x oil immersion lens with a numerical aperture of 1.4. A 488nm laser for imaging GFP. The imaging chamber was maintained at 37°C and 5% CO 2 (Okolab) with a 96 well plate adaptor. Z-stacks were taken every 420nm for 15 μm using an ASI MS-2000 stage controller.