Silencing of Plasma Membrane Ca2+-ATPase Isoforms 2 and 3 Impairs Energy Metabolism in Differentiating PC12 Cells
Figure 8
Determination of βIII-tubulin in differentiated PC12 cell lines. (a) βIII-Tubulin level was determined by immunoblotting in the presence or absence of oligomycin. (b) Densitometric quantification of bands intensity. The results are presented as arbitrary units (AU) obtained after normalization to endogenous GAPDH level. versus control cells; oligomycin treated versus nontreated cells. (c) Representative images of immunofluorescent staining of βIII-tubulin in single cells fixed with paraformaldehyde. The images were taken using TCS SP5 confocal microscope. Scale bar 20 μm. (d) Decrease in βIII-tubulin fluorescent signal following 3 h treatment with oligomycin from randomly selected 10 cells. Fluorescence ratio was calculated as , where is a fluorescence measured following oligomycin treatment and is a corresponding fluorescence intensity in the absence of oligomycin. versus control cells. : mock-transfected PC12, _2: PMCA2-reduced PC12, and _3: PMCA3-reduced PC12.
