Supplementary Figures

Files in this Data Supplement:

• Supplementary Figure S1 - 5.12 MB (.tif)
  RT-PCR analysis of the VSX1 gene in human corneal tissues. VSX1 amplification was not observed using RNAs extracted from 14 corneal tissues obtained from human donors (lanes 1-14). VSX1 expression was found instead in cultured stromal cells grown in the presence of serum, (lane 15). b-actin was used as an internal control.

• Supplementary Figure S2 - 3.1 MB (.tif)
  VSX1 immunostaining in mouse normal and wounded corneas. Scid/beige mice (n=4) were used in this study after pre-operative examination for exclusion criteria such as ocular disease, wounds or infection. The untreated eyes served as normal controls. All animals were cared for and used in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research. Mice were anesthetised with ketamine (10 mg/kg body weigth) / xylazine (0,5 mg/kg body weight) by intraperitoneal injections. Wounds were generated either through alkali burns by using 1-2 drops of 1M NaOH on ocular surface (A) or corneal epithelium removal by means of a rust ring remover (B). The mice were sacrificed 15 days post-healing. Eyes were enucleated immediately after euthanasia and 5- to 7-mm sections obtained from fixed tissues as described in the Materials and Methods. Sections were analysed by indirect immunofluorescence by using VSX1 pAb (1:50, Santa Cruz Biotechnology). A strong VSX1 expression was detected both in corneas wounded through alkali burn treatment (A) and mechanical removal of the epithelium (B), but never in non-wounded controls (C). These observations are consistent with the previously data obtained on wounded human corneas showing that VSX1 is expressed in injured corneal stroma.