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. 2010 Oct 20;10:77. doi: 10.1186/1472-6750-10-77

Figure 1.

Figure 1

(A) - Electrophoretic profile of in vitro synthesized messenger RNAs. Lane M: 0.5-10 kb RNA Ladder (Invitrogen), lane 1: 1 μg of mRNA-Hsp65, 2: 1 μg of EF-1α mRNA. The electrophoresis was performed in 1.5% denaturing agarose gel stained with ethidium bromide. (BP-Base pair, M-Marker). (B) - Detection of the integrity of the mRNA-Hsp65 by RT-PCR. After transfection of HEK 293 cells with the mRNA-Hsp65 cells were maintained in culture for different periods of time, as showed in the figure, in order to analyze the presence of mRNA-Hsp65. Electrophoresis was run in 1.5% agarose gel and stained with ethidium bromide (NC-Negative PCR control, NT-Not transfected, PC-Positive PCR control). (C) - After contact mRNA-Hsp65 for different periods of time, the total cell lysate was subjected to polyacrylamide gel electrophoresis (12.5%) and the bands transferred to nitrocellulose membrane and incubated with anti-Hsp65 for 2 hours. The reaction was revealed with secondary antibody anti-mouse IgG in the presence of DAB. (RC-Recombinant protein Hsp65, NT-Not transfected)