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. Author manuscript; available in PMC: 2015 Jul 15.
Published in final edited form as: Oncogene. 2013 Dec 23;34(3):334–345. doi: 10.1038/onc.2013.552

Figure 8.

Figure 8

Tax interacts with the autophagy molecular complex containing Beclin1 and PI3KC3. (a) Tax binds to BECN1 and PI3KC3. Tax-GST was co-transfected with the autophagy molecules including FLAG-Beclin1, FLAG-PI3KC3 or FLAG-UVRAG in HEK293 cells. 24 hours following transfection, GST pulldown assay was performed. (b) Co-immunoprecipitation of Tax-GFP and BECN1 in Tax-GFP-established, primary human CD4 T cell lines. (c) Tax binds to the domain situated at aa250-300 of BECN1. Tax-HA was co-transfected with GST-tagged BECN1, BECN1ΔN250, BECN1ΔN300 in HEK293 cells, followed with GST pulldown assay. (d) Depletion of BECN1 by lentivirus transduction of BECN1-specific shRNAs in MT-2 cells as examined by anti-BECN1 immunoblot (upper left panel), and Trypan blue exclusion assay to examine cell viability of MT-2 cells transduced with NS- or BECN1-specific shRNAs. * (p < 0.05) and ** (p < 0.01), as determined by Student's t-test, indicate cell viability differences in BECN1 shRNA-transfected cells as compared to NS shRNA-transfected control cells at corresponding time points. (e) PI3KC3 is depleted by PI3KC3-specific shRNA in SLB-1 cells (upper panel), and the cell viability of PI3KC3-depleted SLB-1 cells vs NS shRNA-transduced SLB-1 cells is shown in lower panel.