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. Author manuscript; available in PMC: 2020 Jun 1.
Published in final edited form as: J Neurooncol. 2019 Apr 22;143(2):231–240. doi: 10.1007/s11060-019-03164-5

Figure 4: Overexpression of exogenous MGMT enhances apoptosis induced by alisertib + carboplatin.

Figure 4:

Cell lines were stably transfected with pCMV6 containing human MGMT or empty vector. A. Western blotting was performed to confirm exogenous MGMT expression. Note: a less sensitive ECL substrate was used to obtain these MGMT bands (Pierce ECL Western Blotting Substrate, Thermo 32106) than was used to determine endogenous MGMT expression in Fig 3A (SuperSignal West Femto Maximum Sensitivity Substrate, Thermo 34095). The same ECL substrate was used for β-actin in both cases. B-D. Cells were treated with alisertib (250 nM), carboplatin (10 μM), irinotecan (5 μM), aliserib + carboplatin or alisertib + irinotecan for 24h hr. Cells were stained with Alexa Fluor 594 annexin V conjugate and analyzed with the Countess II FL cell counter. Average fold changes in annexin V binding are presented. *p < 0.05, **p < 0.01; n=3 for U251 and U87, n=2 for GB30.