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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Clin Cancer Res. 2016 Jul 7;22(21):5265–5276. doi: 10.1158/1078-0432.CCR-16-1003

Figure 5. Bortezomib and oHSV therapy enhances NK cell activation.

Figure 5

A-B) U251T3-mCherry+ glioma cells were pretreated with 12nM bortezomib 16 hours prior to 34.5ENVE infection at an MOI of 0.01. Two hours post 34.5ENVE infection, unbound virus was washed out and cells were overlaid with/without primary human NK cells from three independent donors (D1, D2, and D3). Twenty-four hours later, culture supernatants were analyzed for TNFα and IFN-γ gene expression and secretion by Q-PCR (A) and ELISA (B). C) Flow cytometric analysis of NK cell activating ligands (HLA abc, MICA/B, CD155, CD54, CD112, and CD58) on tumor cells after treatment with bortezomib and/or oHSV as indicated. D) Flow cytometric analysis of NK cell activating receptors TRAIL and CD69 as well as intracellular IFNγ after co-culture with treated tumor cells as indicated. E) Western blot analysis of the cleavage of caspases 8, 3, and PARP in treated tumor cells co-cultured with primary NK cells. GAPDH was used as a loading control.