Figure 1: Analysis of the CBAVD phenotype in the patient. (a) The patient showing a complete absence of bilateral vas deferens (red arrow). (b) The sibling of the patient showing normal development of bilateral vas deferens (red arrow). (c) The heat map of RNA expression in the two brothers. SCNN1B and CA12 RNA levels were decreased in the patient compared to his sibling. The patient showed the lower expression of (d) CA12 and (e) SCNN1B according to his sibling by RT-PCR. The results are presented as the mean ± s.d. (n = 3).*P < 0.05 and **P < 0.01 according to Student's t-test. The reduced protein amount of (f) CA12 and (g) SCNN1B was observed in the patient compared to his sibling by western blot. The western blot results represented that the downregulation of CFTR was observed when (h) SCNN1B and (i) CA12 were knocked down in the 293T and A549 cells, respectively. (j) The interactions were observed between CFTR and SCNN1B, or CA12 in 293T and A549 cells, respectively. (k) The increased ubiquitination degradation of CFTR was observed when SCNN1B and CA12 were downregulated in 293T and A549 cells, respectively. CBAVD: congenital bilateral absence of the vas deferens; SCNN1B: sodium channel epithelial 1 beta subunit; CA12: carbonic anhydrase 12; CFTR: cystic fibrosis transmembrane conductance regulator; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; NC: negative control; IB: immunoblotting; RT-PCR: real-time polymerase chain reaction; IP: immunoprecipitation; s.d.: standard deviation; TRDV2: T-cell receptor delta variable 2; ELOVL7: ELOVL fatty acid elongase 7; KLRC4: killer cell lectin-like receptor C4; VSTM2B: V-set and transmembrane domain containing 2B; KLRC4-KLRK1: KLRC4-KLRK1 readthrough; RLN1: relaxin 1; FSTL1: follistatin-like 1; EIF1P6: eukaryotic translation initiation factor 1 pseudogene 6; MAGI2-AS3: MAGI2 antisense RNA 3; TRBV124: T-cell receptor beta variable 12-4; TRAV12: T-cell receptor alpha variable 1-2; PGA5: pepsinogen 5, Group I (pepsinogen A); PGA3: pepsinogen 3, Group I (pepsinogen A); LIPF: lipase F, gastric type; PGC: progastricsin; ATP4A: ATPase, H+/K+ exchanging, gastric, alpha polypeptide; CA8: carbonic anhydrase 8.