XB-IMG-145282
Xenbase Image ID: 145282
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Fig. 3. Shh facilitates processing of Gli2/3 and inhibits Gli2 nuclear localization in spinal neurons. (A) Representative examples of Western blot assays from whole-cell spinal cord homogenates from wild-type and mGli2-GFP–expressing embryos probed with anti-Gli3 and anti-Gli2, respectively. H2B or GAPDH were used as loading controls. Samples were incubated in the absence or presence of 100 nM SAG for 4 h. Gli-FL: Gli full length; GliR: Gli repressor. (B) Shown are representative examples of transverse sections of neural plate and spinal cord (outlined) from wild-type embryos immunostained for Gli2. Dashed squares are areas magnified in adjacent panels. Orthogonal views are shown to demonstrate subcellular localization of Gli2 immunolabeling. (C–E) Dissociated spinal cord cells from mGli2-GFP–expressing 21-hpf embryos were time-lapse imaged every 15 s. (C and D) Representative examples of imaged cells under indicated treatments. Contour of imaged cells and nuclei are indicated with dashed and solid lines, respectively. Color scale bar represents fluorescence intensity increasing from purple to red. Traces represent changes in nuclear (black) and cytosolic (gray) fluorescence for the given examples. (E) Graph shows mean ± SEM percentage of final change in nuclear mGli2-GFP fluorescence intensity upon addition of indicated agents; n ≥ 27 cells per condition; *P < 0.05 compared with control (vehicle only). (Scale bars: 20 μm in B and 10 μm in C.) Image published in: Belgacem YH and Borodinsky LN (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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