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. 2019 Oct 10;105(9):2316–2326. doi: 10.3324/haematol.2019.223024

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Copyright© 2020 Ferrata Storti Foundation

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Figure 5. — Combined RAL-knockdown and blockade of PI3K/Akt or MEK/MAPK signaling. L-363 cells were transfected with shRNA-expression vectors against RALA or RALB, purified next day by selection for strongly transfected cells, and then treated with PD0325901 (1 μM), MK2206 (1 μM) or BYL-719 (2.5 μM). Cell survival was measured by annexin V/PI staining after 2 days (= day 3 post-transfection). Combination of RALA knockdown with PD0325901, BYL-719 or MK-2206 treatment did not further enhance the already strong apoptosis-induction resulting from RALA depletion alone. RALB knockdown in combination with PD0325901 showed only slight additional apoptosis-induction, whereas in combination with BYL-719 or MK-2206 treatment, the rate of cell death was strongly enhanced and matched that achieved by RALA knockdown.