Ethanol induces expression of Sonic hedgehog and its Gli1 transcription factor in murine kidney.

A) Ethanol-induced accumulation of Sonic hedgehog and Gli1 mRNA is dependent on PTAFR. Kidneys of wild-type BL6 or ptafr-/- mice ingesting ethanol or control diets were harvested and mRNA was extracted for qPCR quantitation and analyzed as in Fig 1. SEM (n = 4); *, p<0.05. B) Shh induction by chronic ethanol ingestion is dependent on PTAFR. Kidneys of wild-type BL6 or ptafr-/- mice fed ethanol or their pair-fed controls were harvested and stained by fluorescent immunohistochemistry for Shh and Fluor 568 secondary antibody. C) Gli1 induction by chronic ethanol ingesting is dependent on PTAFR. Sections of kidneys of mice with the stated genotype and diet were stained with anti-Gli1 and Fluor 488-conjugated secondary antibody as in the preceding panel. D) Shh and Gli1 co-localize in the kidneys of ethanol-fed mice. Image overlay of Panels B and C show Shh and Gli1 co-localize, and that the doubly positive cells are positioned to surround vascular lumens as shown by inclusion of a panel reporting DAPI (fluorescing blue) counterstained nuclei.