Figure 4
Assessment of stress/response pathways by qRT-PCR and microscopic imaging.
(a) qRT-PCR analysis of stress-response pathway-related genes. The graphs show levels of mRNA expression in ONC eyes and the non-treated contralateral eyes. The average for the non-treated eyes was set at 1.0. The relative expression is presented for the ONC eyes (N = 6 each). (b) Schematic representation of the retinal areas imaged to quantify EGFP expression in the retinal flatmounts. EGFP-positive cells were counted in four distinct areas of 1.29 mm2 each (one area per retinal quadrant; filled square). (c) Representative images of EGFP-positive cells in the retinal flatmount imaged by conventional microscopy. EGFP-positive cells in the magnified images of the insets are labelled with arrowheads. (d) Quantification of EGFP-positive cells from photographs obtained by microscopic imaging of the retinal flatmounts. The average for the non-treated eyes was set at 1.0 and the relative number of EGFP-positive cells is presented for the ONC eyes (N = 6 each). Data represent means ± S.E.M., **P < 0.05, *P < 0.1. Scale bar: 100 μm. ONC, optic nerve crush injury; No Tx, no treatment. ARE, antioxidant response element; ATF6, ATF6 response element; HRE, hypoxia response element; Mcp-1, Mcp-1 promoter; NF-κB, NF-κB response element; p53, p53 response element; TGF-β, SMAD-binding element.
