Figure 2

PGE2 does not ostensibly affect apoptosis of HT-29 cells. A HT-29 were cultured for varying lengths of time in the absence of serum or in the presence of 10% FCS, 10 nM PGE2 or 10 μM PGE2. At the indicated time points cells were lysed and PARP cleavage was assessed by western blotting. Vinculin levels were determined to ascertain equal protein loading. Staurosporine was administered at 1 μM as a positive control to induce apoptosis. Quantification is shown as the densitometrically-determined ratio of cleaved to non-cleaved PARP levels. Shown here is one representative experiment out of three. B PGE2 does not affect the number of Annexin V positive cells. HT-29 cells were treated with staurosporine, FCS or the indicated PGE2 concentrations followed by AnnexinV-FITC and propidium iodide staining and FACS analysis. The mean percentage of cells gated in the Annexin+/PI- (early apoptotic) and Annexin+/PI+ area (late apoptotic) ± S.E.M. (n = 3) is indicated below each panel.