TUNEL labeling of RPE cells from mice exposed to cigarette smoke for 6 months.

<p>A. TUNEL labeled (red) RPE nuclei are indicated by the arrows. B. Nuclei are stained with DAPI (blue), as labeled by the arrows. C. Merged image of A and B separating TUNEL from DAPI only stained nuclei. D. Brightfield image of the RPE, choroid (Ch) and sclera (S). E. Merged image of a mouse raised in air for 6 months. Arrows point to blue DAPI without red TUNEL labeling. Bar = 15 μm.</p

Transmission electron microscopy of Bruch membrane of mice exposed to air (A) or cigarette smoke (B,C).

<p>A. Mouse exposed to air shows regular basal infoldings (BI) of the RPE and unthickened Bruch membrane (BrM). B. The RPE show loss of basal infoldings. Bruch membrane is thickened due to an outer collagenous layer deposit (OCL). C. The RPE show more severe loss of basal infoldings than in (B). Bruch membrane contains early basal laminar deposits (*), OCL, and choriocapillaris (CC) basement membrane reduplication (arrows). Bar = 500 nm.</p

Immunohistochemistry of 8-OHdG nuclear labeling of the RPE.

<p>8 month C57Bl6 mouse exposed to smoke for 6 months showing A. DAPI labeled nuclei (arrows); B. 8-OHdG labeled RPE nuclei (arrows); C. Merged image of A and B with the Brightfield image showing violet nuclei (arrows); D. Merged image of DAPI and IgG1 control image with Brightfield image overlay. 8 month old C57Bl6 mouse raised in air showing DAPI labeled RPE nuclei in E and 8-OHdG immunostaining in F; G. Merged DAPI and 8-OHdG immunostained image with Brightfield image overlay showing blue nuclei in H. Brightfield image. RPE, retinal pigmented epithelium; Ch, choroid. Bar = 15 μm. Figure shows representative images from N = 10 mice (50 samples/mouse).</p