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. 2015 Jan 9;8:438. doi: 10.3389/fncel.2014.00438

Figure 5.

Figure 5

Overexpression of CHIP reduces apoptosis in hippocampal slices treated with Tunicamycin. Hippocampal slices were infected with rAAV8-GFP or rAAV8-CHIP. After 13 days in vitro, the slices were incubated with Tunicamycin at 80 μg/mL for 24 h. Analysis of chromatin condensation with TO-PRO3 staining (A–D) of slices treated with vehicle (A), Tunicamycin at 80 μg/mL (B); rAAV8-GFP + Tunicamycin at 80 μg/mL (C); rAAV8-CHIP + Tunicamycin at 80 μg/mL (D). 40x Magnification, Scale bar: 20 μm. (E) Quantification of chromatin condensation in treated/infected slices compared to vehicles. P < 0.01, C.I.: 99.9%, N = 3 independent experiments. (F–H) Immunofluorescence for cleaved caspase 3 (in red) after Tunicamycin treatment (G), vehicle (F) and rAAV8-CHIP + Tunicamycin treated (H). 63x Magnification, Scale bar: 20 μm. (I) Quantification of positive cells for cleaved caspase 3. p < 0.01, C.I.: 99.9%, N = 2 independent experiments. (L) TUNEL labeling (green) counterstained with TO-PRO3 (white). (J) Vehicle, (K) Tunicamycin and (L) rAAV8-CHIP + Tunicamycin. 63x Magnification, Scale bar: 20 μm. (M) Quantification of TUNEL positive cells. N = 2 independent experiments ***p < 0.0001.