Chronic ethanol feeding allows Indian hedgehog escape to urine.

A) Chronic ethanol ingestion induces Ihh protein expression in kidney. Ihh expression in kidney was determined by fluorescent immunohistochemistry in mice ingesting a control (left) or ethanol (right) diet. Nuclei were counterstained blue with DAPI mounting media. B) Ihh is present in the urine of ethanol-fed mice. Urine from either pair-fed control animals or that from mice fed a ramped Lieber-deCarli ethanol diet was collected at the end of the feeding trial. Urinary proteins and kidney homogenates were resolved by SDS-PAGE and immunoblotted for Ihh as described in “Methods.” C) Ihh and KIM-1 co-localize in the kidneys of ethanol-fed mice. Fluorescent immunohistochemistry of KIM-1, Ihh, or DAPI nuclear counterstain of renal sections from mice ingesting a ramped ethanol diet or pair-fed a control diet. Image overlay in the right panel of green KIM-1 and red Ihh shows co-expression in yellow.