Genetic Profiling of Monogenic Hypertension

All participants provided venous blood samples for genetic testing. Genomic DNA was extracted from peripheral blood leucocytes using the QIAamp DNA Blood Mini kit (QIAGEN, Hilden, Germany) by standard protocols. Next-generation sequencing of 41 monogenic hypertension-related genes was performed for the proband (Supplementary Table 1). All coding exons were enriched using custom-made SureSelect Target Enrichment System (Agilent Technologies, Inc., Santa Clara, CA). Captured DNA library were sequenced on Illumina Hiseq X Ten according to standard protocol for paired-end 150bp reads. Paired-end reads were aligned to the reference genome (hg19) using the Burrows–Wheeler Aligner and duplicated reads were marked by Picard.^{12 (link)} SNVs and indels were detected by SAMtools, and Annovar^{13 (link)} was used for annotation. A mutation in γ-ENaC was identified by polymerase chain reaction (PCR) using gene-specific primer pairs: SCNN1G: (GenBank accession number NM_001039): forward primer: 5′-CTTGGGAATCAGGGTTCCTGTG-3′, reverse primer: 5′-AAGCAGGCTTTTTGGTCAGAGT-3′.^{14 (link)} The PCR products were sequenced bidirectionally using an ABI Prism 377 DNA sequencer (Applied Biosystems, Foster City, CA).