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. 2011 Oct 26;31(43):15329–15339. doi: 10.1523/JNEUROSCI.2057-11.2011

Figure 3.

Figure 3.

Low level of spontaneous hair cell regeneration following neomycin treatment is augmented by inhibition of γ-secretase: qualitative observations. All images show cultured utricles treated with neomycin for 1 d followed by increasing periods in culture, with 0.5% DMSO or 50 μm DAPT present in media. All images are en face (x, y) views of the SE. A–D, Low-magnification, brightest point projections through the SE from utricles cultured for 3 d (A), 6 d (B), 10 d (C), or 18 d (D) after neomycin treatment (pNeo), with DMSO in the media. An outline of the approximated striolar region is drawn in A. GFP immunolabeling is green and myosin VIIa immunolabeling is red. E, Low-magnification, brightest point projection through the SE cultured with DMSO until 6 d after neomycin. ATOH1 immunolabeling is green and myosin VIIa immunolabeling is red. F, F′, High-magnification slice of the supporting cell nuclear layer, showing labeling for GFP (green), DAPI (blue), and myosin VIIa (red) (F) or DAPI only (F′) in a DMSO-treated culture at 3 d after neomycin. Arrowhead points to GFP+ nucleus. G, H, GFP+ cells in the presumed extrastriolar region (G) or striolar/juxtastriolar region (H) of a DMSO-treated culture at 10 d after neomycin; GFP is green, and myosin VIIa is red. Double-labeled cells have yellow cytoplasm (H). I, I′. High-magnification slice of the hair cell nuclear layer, showing labeling for GFP (green), myosin VIIa (red), and DAPI (blue) (I) or DAPI only (I′) in a DMSO-treated culture at 3 d after neomycin. Arrows point to myosin VIIa+ cells that have GFP+ nuclei. J, High-magnification, brightest point projection through the SE cultured with DMSO until 6 d after neomycin. ATOH1 immunolabeling is green and myosin VIIa immunolabeling is red. K–N, Low-magnification, brightest point projections through the SE from utricles cultured for 3 d (K), 6 d (L), 10 d (M), or 18 d (N) after neomycin treatment, with DAPT present for the whole culture period. GFP immunolabeling is green and myosin VIIa immunolabeling is red. O, Low-magnification, brightest point projection through the SE cultured with DAPT until 6 d after neomycin. ATOH1 immunolabeling is green and myosin VIIa immunolabeling is red. All low-magnification utricle images are oriented in the same manner as shown in Figure 1E. Scale bars: A–E, K–O (in A), 100 μm; F, F′, I, I′, J (in F), 5 μm; G, H, inset in O (in G), 10 μm.