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F. Yang et al.
XL functions as a substrate to directly inhibit caspases before cleavage;14 therefore, we measured caspase 3/7 activity using luminescent assays. Caspase 3/7 activity was increased in cKO fetal liver cells (Figure 6D). Next, we used Q-VD-OPh hydrate, a pan-caspase inhibitor to treat the
Ter119low erythroid cells in order to rescue the impaired definitive erythropoiesis in Tmem30a cKO erythroid cells (Figure 6E).29 Interestingly, the presence of 50 μM Q-VD- OPh partially rescued erythroid cell maturation of Tmem30a-deficient erythroid cells (Figure 6F).
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haematologica | 2019; 104(10)
Figure 4. Tmem30a deletion results in impaired flippase activity. (A and B) Percentages of Annexin V positive cells within each R population were calculat- ed. All values are Mean±Standard Error of Mean (SEM) of four independent repli- cates. (C) NBD-PS fluorescence profiles of fetal liver cells from wild-type or Tmem30a-decifient embryos after 3-minute (min), 10-min or 15-min incubation. (D) NBD-PS in fetal liver cells after 3-15 min of incubation, presented as geomet- ric mean fluorescence intensity. (E) NBD-PS fluorescence profiles in each R pop- ulation of definitive erythropoiesis from control and cKO, assessed after 3-min incubation. (F) NBD-PS in fetal liver erythroid cells from control and cKO, present- ed as fluorescence intensity. Values are presented as Mean±SEM . *P<0.05; **P<0.01; ***P<0.001; with three replicates. N.S.: not significant.