The potentiation of HF-induced cell death by RAPA was also confirmed using Western blot analysis (Fig 4), in which there was an increase in cleaved PARP protein levels in the cultures treated with both HF and RAPA (2.3 ± 0.08) as compared to those treated with either HF alone (1.52 ± 0.06) (HF vs. HF + RAPA: P ˂ 0.0001, two-tailed t-test, n = 3) or RAPA alone (1.12 ± 0.05) (RAPA vs. HF + RAPA: P ˂ 0.0001, two-tailed t-test, n = 3). Here, TRERF1 is linked to hydrops fetalis.