SHH produced by mouse adrenal glands cofractionates with lipoproteins. (A) Left: Schematic representation of the zones and cell types in the mouse adrenal gland. SHH is produced by subset of subcapsular cortical cells, positive for the steroidogenic marker SF1 (magenta). Right: Adrenal gland sections from Shh^GFP mice stained for GFP and DAPI and Gli1^LacZ mice stained for β-galactosidase and SF1. For each staining, a magnification is shown. Scale bar, 300 µm. (B) Experimental scheme for the isolation and differential centrifugation of SHH from mouse adrenal glands. (C) Western blots of adrenal fractions separated by differential centrifugation, probed for SHH or lipoprotein markers (APOA1, APOE). (D and E) Western blot of density gradient fractions from adrenal supernatants at 16,000 g (D) and 150,000 g (E), blotted for SHH and lipoprotein markers (APOA1, APOE). Density of lipoprotein classes is according to Jonas and Phillips (2008). The experiment was repeated three times with similar results. VLDL, very-low-density lipoproteins.