Figure 8

HDAC4 localization pattern in developing murine optic nerve. (A) Low magnification confocal micrograph showing the P30 optic nerve (ON), optic nerve head and retina (R) double-labeled for SOX2 (green) and HDAC4 (red). Optic nerve sections at E16 (B-E), P5 (F-I) and P30 (J-M) were triple-labeled with DAPI (blue), SOX2 (green) and HDAC4 (red). (B, F, J). Overlays of SOX2 and DAPI to aid in orientation of the image. Subsequent panels did not include DAPI to better show the co-localization of the SOX2 and HDAC3 labels. (C, G, K) HDAC4; (D, H, L) double-label of SOX2 and HDAC4. Composite images in E, I and M show z stacks of optic nerve at high magnification; horizontal yellow lines correspond the x axis plane and vertical yellow lines corresponds to the y axis. The x, z axis is shown at the bottom of each panel, while the y, z axis is shown to the right of the panel. Negative controls at each stage showed a lack of immunoreactivity following preabsorption of antibodies with the peptide immunogen (N-P). Abbreviations: R; retina, ON; optic nerve. Scale bars = 50 μm. Scale bar in B applies to B-D, F-H, and J-L. Scale bar in E applies E, I and M.