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. Author manuscript; available in PMC: 2012 Oct 15.
Published in final edited form as: Clin Cancer Res. 2011 Aug 2;17(20):6467–6481. doi: 10.1158/1078-0432.CCR-11-0812

Figure 3.

Figure 3

Short-lived proteins found in autophagosomes isolated from bortezomib-treated cells stimulate antigen-specific T cells. (A) Expression of the short-lived GFP-OVA fusion protein (rGFP-OVA) or the stable GFP-OVA fusion protein (mGFP-OVA) in HEK 293T cells treated with or without bortezomib was measured by flow cytometry (solid line and filled plot respectively). (B) Proliferation of naïve antigen-specific OT-1 cells measured by CFSE dilution. CFSE-labeled naïve T cells from ovalbumin-specific OT-1 TCR transgenic mice were added to an autophagosome-pulsed APC culture for 3–4 days at 37°C. n=2 experiments. (C) Bortezomib-treated HEK 293rGFP-OVA cells were lysed by sonification and autophagosomes were isolated by differential centrifugation as done previously. CFSE-labeled naïve T cells from ovalbumin-specific OT-1 TCR transgenic mice were added to APC pulsed with different quantities of either purified autophagosomes or total cell lysates and co-cultured for 3 days at 37°C. Proliferation of T cells was measured by CFSE dilution as done previously. n=1 experiment.