Mean intensity of green fluorescence in cells in three microscope fields per well and three wells for each condition was used to determine average Htt levels.
The magnitude of the migrated cells was evaluated by counting the migrated cells in 2 random high-power (×100) microscope fields per well under an Axiovert S 100 light microscope (Carl Zeiss Inc., Weimar, Germany) at ×100 magnification.
The cells were fixed with Diff-Quick solution and visualized under an Axiovert S 100 light microscope (Carl Zeiss Inc., Weimar, Germany) in 3 random high-power (×100) microscope fields per well.
After 24 h incubation, the formation of tubes was photographed with a phase contrast microscopy (10× magnification, Olympus Instruments, Inc ., and quantified by counting branch points in five randomly selected microscope fields per well.
The suspension was mixed with 0.4% trypan blue dye for 5 min at 25°C, and the unstained (viable) and stained (nonviable) cells were counted on a hemacytometer within 5 min in five 40× microscope fields per well.
The grayscale value of desmin and myoglobin expression and the myotubes diameters were measured from randomly selected microscope fields from five different wells of Pc-3.1-transfected and Pc-hR-transfected C2C12 myotubes, respectively, and analyzed by image analytical system HPIAS-1000.
At least 4 random microscope fields were analyzed for each sample well and experiments were repeated at least 3 times.
Five nonoverlapping microscope fields (10× objective) were counted in each well.
The wounds were photographed (8 microscopic fields per well at × 4 magnification using inverted light microscope) immediately and after a specified period of treatment.
Adherent cells were counted independently in six random high-power (×100) microscope fields (HPF /well by three observers unaware of the treatments.
The colonies were count in 10 randomly chosen microscope fields.
