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. Author manuscript; available in PMC: 2019 Aug 14.
Published in final edited form as: Clin Cancer Res. 2012 May 1;18(13):3603–3615. doi: 10.1158/1078-0432.CCR-11-3321

Figure 5. The combination effects of MK-2206 and rapamycin in vitro and in vivo.

Figure 5.

AS, BE2, NGP and SY5Y cells were used. A. NB cells were treated with rapamycin for 48 hrs and cell proliferation was evaluated. Bar, SD. B: NB cells were treated with rapamycin (10nM) for 2 hrs followed by MK-2206 (7.5μM) treatment for another 2 hrs, either alone or in combination. Total protein was extracted to detect P-S6, T-S6, P-Akt, T-Akt and GAPDH levels. C and D: NB cells were treated with rapamycin (10nM) for 2 hrs followed by MK-2206 (7.5μM) treatment and cell proliferation was evaluated after 48 hrs (C) or caspase 3/7 activity was detected after 24hrs (D). Bar, SD. # P<0.05, ##P<0.01: MK-2206/rapamycin-treated cells versus MK-2206-treated cells; ** P<0.05, **P<0.01: MK-2206/rapamycin-treated cells versus rapamycin-treated cells. NS: no statistical significance. E: NGP, BE2, AS and SY5Y cells were pretreated with NAC (100mM) for 2 hrs followed by MK-2206 (7.5μM) and Rapamycin (10nM) treatment, either alone or in combination for 48 hrs. Cell proliferation was detected. Bar, SD. # P<0.05: NAC/MK-2206-treated cells versus MK-2206-treated cells; @ P<0.05: NAC /rapamycin-treated cells versus rapamycin-treated cells, *P<0.05, ***P<0.001: NAC /rapamycin/MK-2206-treated cells versus rapamycin/MK-2206-treated cells. F: Mice bearing NGP tumors were treated with rapamycin at 2.5mg/kg/day or 5mg/kg/day for 2 weeks by IP injection. Total proteins from tumor tissues were extracted for evaluation of P-S6, T-S6, P-Akt and T-Akt. G and H: Mice bearing NGP tumors were treated with MK-2206 (200mg/kg) and rapamycin (5mg/kg) either alone or in combination. Tumor volumes from control and treated groups were compared when the last mouse in control group was euthanized (G). Bar, SE. Mice survivals were compared with the Kaplan-Meier curves (H). ## P<0.01: rapamycin/MK-2206-treated group versus MK-2206-treated group. *P<0.05: rapamycin/MK-2206-treated group versus rapamycin-treated group. I: NB tumors were harvested at end points, total protein was extracted for analysis of P-Akt, T-Akt, P-S6, T-S6 and GAPDH.