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. 2014 Apr 21;9(4):e95977. doi: 10.1371/journal.pone.0095977

Figure 5. Quantification of mRNA and protein expression of TAP1 and MHC class I molecule during T. cruzi infection.

Figure 5

(A) The abundance of TAP1, β2M and HLA mRNAs were determined by real-time RT-qPCR using the total RNA from HeLa cells treated with IFN-γ and/or infected with T. cruzi. The relative expression of the transcripts was calculated by normalization with GAPDH and HPRT1 housekeeping genes using the 2−ΔΔCt method. The mRNA levels were plotted relatively to “IFN-γ” experimental condition (HeLa treated 24 h with IFN-γ). Each value represents the mean ± standard deviation of three independent experiments. (B) Lysates (25–50 µg) of HeLa cells treated with IFN-γ and/or infected with T. cruzi were analyzed by western blot using human anti-TAP1 and anti-MHC class I antibodies as indicated. Infection was confirmed using anti-tubulin antibody. (C) Protein levels were determined by densitometry and plotted using the expression of α6 subunit as experimental normalizer. Each value represents the mean ± standard deviation of three independent experiments.