Figure 5.—

The transient transfection of D.Mel-2 cells with the Dmel\TIP60/RNAi construct results in deleterious effects on cell growth and reduction of endogenous Dmel\TIP60 transcript levels. (A–D) D.Mel-2 cells visualized at ×200 magnification using phase/contrast optics. (A) Cells transiently transfected with pAc5.1/V-5-His/LacZ (unstained). (B) The same cells as in A stained with X-Gal showing transfection efficiency at 77%. (C) Cells transiently transfected with Dmel\TIP60/control construct, shown 24 hr post-transfection. (D) Cells transiently transfected with Dmel\TIP60/RNAi construct, shown 24 hr post-transfection. Arrows point to morphologically defective cells. (E) Semiquantitative RT–PCR analysis of Dmel\TIP60 and RP49 transcript levels. RNA was isolated from cells (shown above) 24 hr post-transfection. Equal amounts of RNA for each sample were subjected to cDNA preparation using RT priming with random hexamers and PCR using primer sets specific for Dmel\TIP60 that did not amplify RNAi target sequences and primer sets specific for RP49 internal control. All experiments shown were repeated at least three independent times with consistent results.