Similar sections were stained with the anti TIMP 1 and anti TIMP 3 anti-bodies to find cells synthesising these proteins. As indicated in Fig. 4, these cultures exhibited the greatest amount of TUNEL positive cells and the extent of TIMP 3 induced apoptosis was considerably paid down in the cultures co infected with RAdTIMP 1 o-r pre incubated with rTIMP 1. Representative pictures representing the reduction in the amount of TUNEL and caspase 3 stained cells in cultures company infected with RAdTIMP 3 and RAdTIMP 1 are shown in Fig. 5aef. TUNEL staining was used to visualise apoptotic cells in sections of standard, non Ivacaftor clinical trial scarred keratoconic and scarred keratoconic corneas. A photograph of a sectioned scarred keratoconic cornea shows several TUNEL positive cells in the epithelium of the corneas but nothing within the posterior stroma. A lot more were found in the anterior stroma of the scarred keratoconic corneas, although relatively few numbers were found in the anterior stroma of the standard and non scarred keratoconic corneas. Over all, while all epithelial cells of normal and keratoconic corneas reacted strongly with the antiTIMP 1 antibody, relatively several TIMP 1 creating stromal cells were discovered, especially in the normal corneal parts. Those who were contained in the keratoconic corneal sections were located primarily in the sections of scarred keratoconic corneas and most abundantly in the anterior stroma, just below Bowmans membrane. As opposed to the design of TIMP 1 staining in the corneal epithelium, only the basal cell layer Cellular differentiation did actually respond with the antiTIMP 3 antibody. The distribution of the TIMP 3 positive stromal cells, which was most abundantly found in the anterior stroma of scarred keratoconic corneas, was but much like that of the TIMP 1 positive stromal cells. In-addition, since TIMP 3 is just a matrix related protein, basic staining was observed through the stroma of all corneas which were examined. No TIMP 3 staining was seen in the Bowmans layer, Descemets membrane, the endothelium or control sections incubated with control IgG serum or without primary antibody. The results of previous work suggested that in stromal cell cultures based on scarred keratoconic corneas TIMP 1 production was up managed and TIMP 3 may be solubilised when its matrix natural compound library binding proteins were degraded by proteolytic enzymes. Hence, although little of-the TIMP 3 synthesised by the transfected stromal cells of normal corneas was recovered in a soluble form in their lifestyle media, the TIMP 1 and TIMP 3 contents of the soluble protein extracts of normal and keratoconic corneas were tested by ELISA.