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For testing the expression level of GUS in leaves, fully expanded 4th leaf of each seedling at four-leaf stage was used.
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qRT-PCR was carried out to test the expression level of genes around the predicted insertion point.
We selected a number of genes that had roles in transcriptional regulation, development and signal transduction and tested the expression level of these genes using quantitative RT-PCR.
Since previous yeast two hybrid results indicated possible interaction of SNAP29 with other members of the EHD family [12], we also tested the expression level and intracellular localization of other EHDs in the absence of SNAP29.
Therefore, we tested the expression level of miR-29a using the hepatoma cell lines which were reported with different metastasis potential as a model, such as MHCC-97H and MHCC-97L [24].
To further confirm that hsf1-R206S, F256S cells had enhanced basal activation of Hsf1 at 25°C, we tested the expression level of numerous known transcriptional targets of Hsf1 in these cells.
We also tested the expression level of NRGN in these samples by performing qRT PCR.
Since cross-talk between EGFR and IGF-1R exists we tested the expression level of IGF-1R in these lines.
Using these cells, we analyzed the changes in the expression of genes and proteins, tested the expression level of miR-9, and applied a target validation system.
We then tested the expression level of ZEB2 in 293T cells transfected with empty vector, the pSG5-miR-142-wt pSG5-miR-142-wt pSG5-miR-142-wt pSG5-miR-142-wt
Since p53 is involved in cell growth and apoptosis, we tested the expression level of p53 in DHODH-knockdown cells; no differences were found among the groups.
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