Figure 1

A: Dual label immunofluorescent laser scanning confocal microscopy of ocular surface tissue sections from C57BL/6 mice for interleukin 2 receptor alpha (CD25, green) and beta chains (CD122, blue) with propidium iodide (red) nuclear counterstaining in nonstressed controls (NS), 5 days (D) of desiccating stress (DS5) and DS5 treated with topical doxycycline (DS5+Doxy) in C57BL/6 mice. A turquoise color indicates co-localization of both markers. Note partial disappearance of CD25 with preservation of CD122 after DS5 (arrows) in the conjunctival epithelia. Scale bar = 50 μm 1. B. Tissue sections prepared for in situ zymography (in situ Z) in nonstressed controls (NS), 5 days (D) of desiccating stress (DS5) and DS5 treated with topical doxycycline (DS5+Doxy) in C57BL/6 mice. Scale bar = 100 μm. 1. C. Merged images of laser scanning confocal fluorescent microscopy of ocular surface tissue sections stained for matrix metalloproteinase 9 (MMP-9, in green) with propidium iodide (PI, red) nuclear counterstaining in NS controls, DS5 and DS5+Doxy groups in C57BL/6 mice. Scale bar = 50 μm 1. D. Representative Western blot showing effect of DS on CD25 expression in corneal (CO) and conjunctival epithelial (CJ) lysates. 1. E. Bar graphs are mean + standard error mean of CD25 band intensities in 3 independent Western blots (arbitrary units).