The existence of predicted genes was verified by RT-PCR analyses designed to amplify spliced sequences.
The down-regulation of these genes was verified using qPCR, and downstream effects were further tested using NR activity assays.
As a result, the presence or absence of 182 genes out of the 221 genes was verified.
Overexpression of candidate genes was verified by Western blotting.
Regulation of subsets of these genes was verified by quantitative PCR in an independent experiment.
Promoter methylation status of these genes was verified by bisulfite genomic sequencing of CpG islands.
Confirmation of the differential expression of genes was verified and shown in Figure 4.
The deletion of the genes was verified by Southern analysis (see Figure S2).
The mRNA expression for several novel glomerular-enriched genes was verified by qRT-PCR.
The function of these genes was verified using GeneDeks V3 software (http://www.genecards.org/ [55]- Table S4).
The expression of all but one of the newly discovered genes was verified by RT-PCR.
