−254C>G SNP in the TRPC6 Gene Promoter Influences Its Expression via Interaction with the NF-κB Subunit RELA in Steroid-Resistant Nephrotic Syndrome Children

Figure 1

Effect of the −254C>G SNP on TRPC6 promoter activation in podocytes and HEK 293T cells. (a) Schematic diagram of a series of plasmids used for the promoter assay. 1332 bp DNAs from the TRPC6 promoter region containing the −254C or G allele were subcloned into luciferase expression plasmids (shown as LUC1, LUC2, and LUC3), and the control was a sequence without a −254 site. The transcription start site is denoted by +1 and indicated by a horizontal arrow. (b and c) The −254G allele (LUC1/2/3-M) significantly enhanced the TRPC6 promoter activation. Podocytes (b) and HEK 293T cells (c) were transfected with the different pGL3 plasmids (LUC1-W/M, LUC2-W/M, LUC3-W/M, control, and pGL3-basic), and the luciferase activities were determined in the cell transfection studies. The results were expressed as the ratio of firefly to Renilla luciferase activity of quadruplicate cultures, representative of three experiments. ; luciferase activity of LUC1/2/3-M compared with that of LUC1/2/3-W, respectively.

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