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. 2017 May 5;8:15182. doi: 10.1038/ncomms15182

Figure 2. Autophagy in the developing neuroepithelia of normal and NTD embryos.

Figure 2

(a) indicators of histological sectioning plane for b. Red lines indicate the levels of sections shown below. For each embryo, five sections from top to bottom were chosen to detect autophagy levels, forebrain, midbrain, hindbrain and spinal cord were indicated in each picture. Scale bars: 300 μm. (b) representative images showing autophagy levels (green LC3-GFP puncta) in sections of the forebrain, midbrain, hindbrain and spinal cord. E10.5 embryos from nondiabetic (ND) and diabetic mellitus (DM) females mated with GFP-LC3 transgenic males were used for autophagy examination. Scale bars: 300 μm. Individual green LC3-GFP puncta can be clearly visualized in high magnification images in Fig. 4. (c) quantification of relative autophagy levels by fluorescence intensity using the ImageJ software in midbrain and forebrain (three serial sections of each area per embryo, and three embryos from three dams were analysed) in the corresponding brain regions. ND, nondiabetic embryos; DM, diabetic mellitus embryos; NTD, exencephaly. *indicates significant difference (P<0.05) compared with the ND groups in one-way ANOVA followed by Tukey tests.