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Experiments were repeated in 3 different passages of cell line.
The process was repeated three times using different passages of cell lines to ensure reproducibility.
These findings may be explained by the speculation that such tumours expressed increased levels of IGF2 mRNA at the critical time of tumorigenesis, but not at the time of surgical resection or after many passages of cell culture.
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Cell passage stability was evaluated with 2 independent runs; each run included 2 passages of cells with the same NC and PC preparations on the same plate, 1 passage ≤ 10 (passages 9 and 10) and the other ≥ 20 (passages 21 and 22).
Differentiations were repeated with three successive passages of cells for a total of three grows.
Experiments were repeated at least 2-3 times using different passages of cells on different days.
This observation was reproducible across three separate passages of cells (Additional file 1, Figure S4).
3rd and 4th passages of cells were used for tumor experiments.
The experiments were repeated at least 3 times using different passages of cells on different days.
The third or fourth passages of cells were used for experiments.
RNA fractions were prepared from four different passages of cells that were approximately 80% confluent, to provide some biological variability.
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