Computational protocol: Histone deacetylase 3 indirectly modulates tubulin acetylation

Similar protocols

Protocol publication

[…] Cells were grown on glass coverslips, fixed with 2% paraformaldehyde in PBS and stained using standard procedures []. Cells were imaged using a Deltavision Deconvolution microscope or a Zeiss 880 Confocal with AiryScan (). Images used for quantification () were captured using the Deltavision system, using the same exposure settings. The fluorescence intensity of a 75 pixels×75 pixels square was measured using ImageJ (NIH), for nine cells, from at least two separate experiments. Images for mitotic cells were captured using the Deconvolution microscope using the ×100 objective, 1×1 binning and a stack size of 20 over a total of 4 μm. The size of the spindles and the area was measured from the final maximum intensity projection image for the total stack, using ImageJ. [...] To visualize microtubule dynamics in live cells, cells were infected with an adenovirus for eGFP–end-binding 1 (EB1), and cells were imaged by time-lapse imaging on a Deltavision deconvolution microscope, fitted with an incubator set to a temperature of 37°C, and using the ×63 oil objective. Prior to imaging, cells were treated for 1 h with increasing concentrations of MI192. Images of eGFP–EB1-labelled microtubule tips were taken every 2 s, and movement of the EB1 fluorescent spots were subsequently tracked using the MTrackJ plug-in in ImageJ. […]

Pipeline specifications

Software tools ImageJ, MTrackJ
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Homo sapiens
Diseases Prostatic Neoplasms