Journal: Scientific Reports

Article Title: FTY720 Induces Autophagy-Associated Apoptosis in Human Oral Squamous Carcinoma Cells, in Part, through a Reactive Oxygen Species/Mcl-1-Dependent Mechanism

doi: 10.1038/s41598-017-06047-9

Figure Lengend Snippet: ( A ) Dose-dependent effects of FTY720 on the phosphorylation of Akt, mTOR, GSK3β, and IKKα/β, and the expression of NF-κB, and survivin in SCC2095 cells. Cells were treated with FTY720 in 5% FBS-supplemented DMEM/F12 medium for 24 h, and cell lysates were immunoblotted as described in Material and Methods. ( B ) Effect of FTY720 on the nuclear translocation of NF-κB. SCC2095 cells were treated with 5 μM FTY720 for 24 h, stained with anti-NF-κB, and examined by confocal microscopy. ( C ) Dose-dependent effects of FTY720 on the expression of Bcl-2, Bcl-xL, Mcl-1, Bax, and Bak. ( D ) Time-dependent effects of FTY720 on the expression of Mcl-1. ( E ) Mcl-1 mRNA expression measured by RT-PCR in cells treated with FTY720. ( F ) Effect of proteasome inhibitor on Mcl-1 expression. Cells were exposed to FTY720 in the presence or absence of 200 nM MG132 for 24 h. ( G ) Effect of ectopic Mcl-1 expression on apoptotic related proteins. SCC2095 cells were transfected with control vector or Mcl-1 plasmid for 24 h and then exposed to FTY720 for 24 h. Whole cell extracts were subjected to Western blot analysis. ( H ) Effect of Mcl-1 overexpression on the viability of SCC2095 treated with FTY720 (7.5 μM) for 24 h. After incubation, cells were analyzed using flow cytometry. Columns , means; bar , S.D. * P < 0.05.

Article Snippet: Mcl-1 plasmid was obtained from OriGene Technologies, Inc. (Rockville, MD).

Techniques: Expressing, Translocation Assay, Staining, Confocal Microscopy, Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Western Blot, Over Expression, Incubation, Flow Cytometry