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. Author manuscript; available in PMC: 2015 Apr 15.
Published in final edited form as: J Immunol. 2014 Mar 17;192(8):3816–3827. doi: 10.4049/jimmunol.1301738

Table II.

Populations of subretinal cells 7 and 21 days after light exposure at 10,000 lux for 30 min

mouse models days after light Iba-1-positive (%) NIMP-R14-positive3 (%) CD3-positive (%) counted cells1/8 slides
Abca4-/-Rdh8-/- 7 days 97.9 ± 0.9 2.1 ± 0.9 0 191.5 ± 13.5
Abca4-/-Rdh8-/- 21 days 90.3 ± 4.2 6.9 ± 1.0 2.4 ± 2.2 59.7 ± 18.6
Ccl3-/-Abca4-/-Rdh8-/- 7 days 98.6± 2.8 1.3 ± 0.5 0 198.2± 21.7
Ccl3-/-Abca4-/-Rdh8-/- 21 days 91.4 ± 6.2 5.4 ± 0.7 2.8± 1.5 147.7 ± 24.1
GFP-positive (%) counted cells2
Cx3Cr1gfp/ΔAbca4-/-Rdh8-/- 7 days 100 1034
1

Cryosections were prepared from every 200 μm distance from the edge to edge (8 slides/eye), and IHC was performed with anti-Iba-1 Ab for microglia/macrophage, anti-Nimp-R14 Ab for neutrophil and anti-CD3 Ab for T cell. Numbers of subretinal cells were counted from these sections, and the ratio of these cells was calculated.

2

Flat-mount eyes were prepared and subretinal cells with GFP and autofluorescent signals were counted under fluorescent microscope.

3

NIMP-R14 recognizes Ly6C in addition to Ly6G, and can therefore react with activated microglia/macrophages. As shown in supplemental Fig S3, subretinal cells with stronger signals over background or weakly stained cells were counted as positive cells.