Figure 10.
Apoptosis is enhanced in AP-1 cells expressing L188P, G383D, E547*, and W570* mutants. Measurement of apoptosis by flow cytometry analysis of AP-1 cells transfected with NHE6GFP WT and CS-linked variants is shown. Forty eight hours post-transfection, cells were labeled with annexin V–APC and PI, and 1 × 104 GFP-positive cells were examined by flow cytometry for each transfectant. A, annexin V and PI double-negative cells represent viable cells. B, PI-positive cells represent necrotic cells. C, annexin V–positive cells represent early apoptotic cells. D, annexin V and PI double-positive cells represent late apoptotic cells. Results are shown as mean ± S.D. of three independent experiments. Significance was determined by one-way ANOVA (A, F value = 27.7, p value = 8.5 × 10−8; B, F value = 2.1, p value = 0.11; C, F value = 40.9, p value = 4.9 × 10−9; D, F value = 5.4, p value = 0.0025), with a post hoc Tukey test; ★, p < 0.05; ★★, p < 0.01; and ★★★, p < 0.001.