Journal: Oncotarget

Article Title: A c-Jun N-terminal kinase inhibitor, JNK-IN-8, sensitizes triple negative breast cancer cells to lapatinib

doi: 10.18632/oncotarget.20581

Figure Lengend Snippet: (A) Cells were treated with 5μM JNK-IN-8 (JIN) and/or 3μM lapatinib (Lap) for MDA-MB-231 cells, and 4μM JIN and/or 7μM Lap for MDA-MB-436 cells for various time points. Cells were also transfected with NFκB luciferase reporter plasmids. Line graph points represent normalized luciferase values from 3 technical replicates. (B) MDA-MB-231 cells were treated with either vehicle (DMSO), 5μM JIN and/or 3μM Lap for 48 hours in full media. Whole cell lysates were probed for phospho-p65(ser536), p65, p50, IKKβ, and IKKα. (C) MDA-MB-231 cells were treated with 5μM JIN and/or 3μM Lap for various time points and transfected with a luciferase reporter plasmid under the control of the NQO1 promoter containing Nrf2/ antioxidant response elements (ARE). Line graph points represent normalized luciferase values from 3 technical replicates. (D) Relative levels of glutathione were measured in MDA-MB-231 and MDA-MB-436 cells after 72 hours of treatment in DMSO, JIN, Lap, or JNK-IN-8 and lapatinib (J+L). (E) Levels of NADP + and NADPH were measured in MDA-MB-231 and MDA-MB-436 cells after 72 hours of treatment in DMSO, JIN, Lap, or J+L. (F) MDA-MB-231 cells were non-transfected, transfected with empty vector (EV), or expression plasmids for p65 (p65 OE) and Nrf2 (Nrf2 OE). Twenty four hours after transfection cells were treated with either vehicle (DMSO), 5μM JIN and/or 3μM Lap for 72 hours in full media. Cell viability was assayed using MTS.

Article Snippet: The next day cells were transfected with 200ng (96 well) or 2μg (6 well) of empty vector (pcDNA.3), p65 (RelA cFlag pcDNA3 was a gift from Stephen Smale (Addgene plasmid # 20012)), or Nrf2 (pCDNA3-Myc3-Nrf2 was a gift from Yue Xiong (Addgene plasmid # 21555)).

Techniques: Transfection, Luciferase, Plasmid Preparation, Control, Expressing