Fig. 1
From: Macrophage-Derived Heme-Oxygenase-1: Expression, Regulation, and Possible Functions in Skin Repair
Wound repair is characterized by strongly increased HO-1 mRNA and protein levels. The complete healing process (A), or the first 24 hr of repair (B) were analyzed for HO-1 mRNA and protein expression. Total cellular RNA (20 µg) from normal and wounded back skin of Balb/c mice was analyzed by RNase protection assay for expression of HO-1 (A and B, upper panels). Sixteen wounds (n = 16) from the backs of four animals were excised for each experimental time point and used for RNA isolation. The time after injury is indicated at the top of each lane. “Control skin” refers to non-wounded skin. One thousand cpm of the hybridization probe were added to the lane labeled “probe.” Expression of GAPDH mRNA is shown as a loading control. The degree of HO-1 induction as assessed by PhosphoImager (Fuji) analysis of the radiolabeled gels is shown schematically in the middle panels. Note, that every experimental time point represents a total of 32 wounds from two independent animal experiments. Total protein (50 µg) from lysates of nonwounded and wounded back skin (10 hr, 16 hr, and days 1, 3, 5, 7, and 13 after injury, indicated at the top of each lane) were analyzed by immunoblotting for the presence of HO-1-specific protein. Eight wounds from the backs of four animals were excised for each experimental time point and used for protein isolation. “Control skin” refers to nonwounded skin. HO-1 protein is indicated by an arrow.