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Fig. 2 | BMC Medical Genetics

Fig. 2

From: Functional characterization of two enhancers located downstream FOXP2

Fig. 2

Molecular characterization of FOXP2-Eproximal and FOXP2-Edistal. a. PCR analysis. Two oligos flanking the deleted regions were used to amplify the genomic DNA from two FOXP2-Eproximal and two FOXP2-Edistal deleted representative SK-N-MC and HEK293 cells. Black triangles show the size of the PCR products. b. Representative Sanger sequencing chromatogram showing the sequences of the junctions of the FOXP2-Eproximal (top) and FOXP2-Edistal (bottom) genomic deleted regions in a SK-N-MC cells. c. Western blot analysis of cell lysates of SK-N-MC with FOXP2-Eproximal deleted, with FOXP2-Edistal deleted, and of SK-N-MC control cells electroporated with pLV-U6#H1#-C9G plasmid for FOXP2 (top) or MDFIC (bottom) proteins analysis. d. qRT-PCR analysis in triplicate as technical replicates of six SK-N-MC cell clones with FOXP2-Eproximal or FOXP2-Edistal deletions, control cells are SK-N-MC cells electroporated with the pLV-U6#H1#-C9G plasmid. Both deleted and control values are normalized to those of the internal reference gene hGUSB. Levels of expression of FOXP2 (up) and MDFIC (down) are represented by the fold change relative to that of empty vector control cell line, which was normalized to 1. Data from three or more independent experiments were analysed by two-tailed unpaired t-test. NS, non-significant; * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001

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