Figure 4
Platelet extract antagonism of drug-mediated inhibition of apoptosis. Apoptosis assays. On the left are shown examples of the results obtained using the Muse Annexin V kit (A) or Caspase-3/7 kit (B) to evaluate the percentage of apoptotic PLC/PRF/5 Hep3B cells treated with 2.5 μM of Regorafenib (A-B) or Sorafenib (A) and cultured whit 3.75 × 107 hPL or equivalent FBS are expressed in the relative graph as the mean of three independent experiments. On the right the mean of three independent experiments is plotted in the relative graph. The results are expressed as mean ± SD. ***p < 0.0001. The anti-apoptotic effect of hPL was also evaluated at the molecular level (C). The representative Western blot shows Hep3B cells treated with 2.5 μM Sorafenib with and without hPL from 3.75 × 107 platelets. hPL caused increased levels of anti-apoptotic factors (survivin, Bcl-xL, P-AKT) and decreased levels of pro-apoptotic (Bax, Bim) factors.