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. 2012 Dec 18;7(12):e51804. doi: 10.1371/journal.pone.0051804

Figure 1. Delta-amastin is more abundant in less infective T. cruzi extracellular amastigotes. A.

Figure 1

mRNAs in EAs of the G strain are less abundant when compared to EAs of the CL-Brener clone T. cruzi . Transcript levels were determined by quantitative real-time PCR using SYBR® Green I chemistry. qRT-PCR was performed on RNA samples from EAs of G and CL strains. The comparative mRNA levels were determined after normalization with GAPDH amplicons. Standard deviations are derived from three replicates. *p<0.05 B. mRNA corresponding to amastin is preferentially expressed in amastigotes from CL Brener clone. Northern blot analyses of total RNA (10 µg) from T. cruzi epimastigotes (E), trypomastigotes (T) and amastigotes (A) from CL-Brener clone or the G T. cruzi strain was submitted to electrophoresis and blotted on nylon membranes by standard procedures. Each blot was hybridized with amastin probe previously labeled with [α-32P]-dCTP. To determine equal loading of RNA, the 1.2% agarose/MOPS/formaldehyde gel was stained with ethidium bromide (bottom panel).