Mutation of hepatocyte nuclear factor–1β inhibits Pkhd1 gene expression
and produces renal cysts in mice
J. Clin. Invest. Thomas Hiesberger, et al. 113:814 doi:10.1172/JCI20083 [
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Figure 2DNase hypersensitive site mapping and deletion analysis of the mouse
Pkhd1 promoter. (A) Structure of the 5′ end of the
Pkhd1 gene. Boxes indicate exons. Bent arrow indicates the transcription initiation site at +1. Bar indicates the 3′ probe used for indirect end labeling. Vertical arrows indicate hypersensitive sites. (B) Southern blot of genomic DNA from mIMCD-3 cells (right) and 10T1/2 cells (left) after digestion with graded concentrations of DNase I. Open arrow indicates the parental 8.1-kb
EcoRI fragment. Closed arrows indicate sub-bands corresponding to hypersensitive sites located at the positions indicated on the right. (C) Northern blot showing endogenous expression of
Pkhd1 (upper panel) and HNF-1β (middle panel) in mIMCD-3 cells (lane 2) and absence of expression in 10T1/2 cells (lane 1). Lower panel shows expression of GAPDH as a loading control. (D) Deletion analysis of the
Pkhd1 promoter. Left panel shows plasmids containing fragments of the
Pkhd1 promoter linked to a promoterless luciferase (Luc) reporter gene. Bent arrow indicates the transcription initiation site at +1, gray boxes indicate exons, and black boxes indicate the consensus HNF-1 site. Right panel shows luciferase activity in transfected mIMCD-3 cells (white bars) and 10T1/2 cells (gray bars). Data are presented as mean ± SE of six to nine independent transfections. *
P < 0.05 compared with empty pGL3-Basic.
