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. Author manuscript; available in PMC: 2012 Apr 10.

Published in final edited form as: Virology. 2011 Feb 15;412(2):357–365. doi: 10.1016/j.virol.2011.01.026

Fig. 3 — The CR1 region of LANA1 inhibits MHC I peptide antigen presentation in cis. (A) Representative results from flow cytometry analysis for SIINFEKL presentation on 293KbC2 cells expressing EGFP-LANA1_OVA constructs (Fig. 2A). Cells were stained with APC anti-mouse MHC class I Kb-SIINFEKL (25-D1.16) 24 h after transfection. Samples were gated for EGFP positivity to ensure construct expression and then the percentage SIINFEKL presenting cells were determined by APC positivity. All experiments were repeated at least three times using either APC or Phycoerythrin (PE) 25-D1.16 (eBioscience) for detection and show similar results. (B) Quantitation of SIINFEKL presentation from three independent experiments (mean±S.D.). P=0.0001 (EGFP and EGFP_OVA), 0.0023 (EGFP_OVA and LANA_OVA), 0.0049 (LANA_OVA and LANAΔCR1_OVA), 0.0135 (LANA_OVA and LANAΔCR123_OVA), 0.0311 (LANA_OVA and LANAΔCR12_OVA). P values between different groups, were obtained by two-tailed Student's t test using Prism software (GraphPad). (C, D) The effect of different pretreatment conditions on antigen presentation. Cells were pretreated with MG132 (proteasome inhibitor) (C), interferon gamma (immunoproteasome inducer) (D) and then SIINFEKL peptide presentation was determined by flow cytometry analysis.

Fig. 3 — The CR1 region of LANA1 inhibits MHC I peptide antigen presentation in cis. (A) Representative results from flow cytometry analysis for SIINFEKL presentation on 293KbC2 cells expressing EGFP-LANA1_OVA constructs (Fig. 2A). Cells were stained with APC anti-mouse MHC class I Kb-SIINFEKL (25-D1.16) 24 h after transfection. Samples were gated for EGFP positivity to ensure construct expression and then the percentage SIINFEKL presenting cells were determined by APC positivity. All experiments were repeated at least three times using either APC or Phycoerythrin (PE) 25-D1.16 (eBioscience) for detection and show similar results. (B) Quantitation of SIINFEKL presentation from three independent experiments (mean±S.D.). P=0.0001 (EGFP and EGFP_OVA), 0.0023 (EGFP_OVA and LANA_OVA), 0.0049 (LANA_OVA and LANAΔCR1_OVA), 0.0135 (LANA_OVA and LANAΔCR123_OVA), 0.0311 (LANA_OVA and LANAΔCR12_OVA). P values between different groups, were obtained by two-tailed Student's t test using Prism software (GraphPad). (C, D) The effect of different pretreatment conditions on antigen presentation. Cells were pretreated with MG132 (proteasome inhibitor) (C), interferon gamma (immunoproteasome inducer) (D) and then SIINFEKL peptide presentation was determined by flow cytometry analysis.