Fig. 2.

Lipid scramblase TMEM16F causes transient and reversible PS exposure on NK cells. a Expression of human TMEM16F-encoding RNA (Ano6) in YT-S cells expressing GFP alone or in combination with wild-type TMEM16F or the aspartate 408-to-glycine 408 (D408G) TMEM16F mutant was measured by RT-PCR. 18S RNA was used as control. b, c Annexin V and PI staining was performed on YT-S cells expressing GFP alone or in combination with wild-type TMEM16F or D408G TMEM16F that were treated or not for 5 min with the calcium ionophore ionomycin [10 μM; in ethanol (EtOH)] or A23187 (10 μM in EtOH). Representative dot plots are shown in (b), while the statistics for seven independent experiments are shown in (c). d Same as (b), except that the cells were incubated or not with BAPTA-AM (100 μM). The data are representative of two independent experiments. e Same as (b), except that ionomycin was washed out, and the cells were incubated for the indicated periods at 37 °C in the culture medium. Representative of two independent experiments. NS not significant; *p < 0.05; **p < 0.01; ***p < 0.001 (two-tailed Student’s t tests). The data are presented as the means ± s.e.m