Why? What is the specific question or hypothesis being tested for this figure? H

Question

Why? What is the specific question or hypothesis being tested for this figure?

How? Methods: Describe how the experiment was performed.

What? Results. Summarize the experimental results, highlighting the relevant controls and main conclusions that can be made only from the data in this figure.

So what? Conclusions. Summarize the conclusions from this figure (and only this figure!)

Fig. 3. Cdh6 knockdown reduces NCC detachment and EMT. (A) NCC EMT behaviors and imaging region. (B-D) Time-lapse images (dorsal views, anterior left, confocal z-projections, beginning at 14 hpf) of NCCs expressing GFP-CAAX. First frames show a single DIC plane for reference. Yellow dashed lines mark basal neuroepithelial surfaces and white dashed lines mark NCC Membrane 2/3. Blebbing Basal (C,D) Cdh6 knockdown with coh6SpIMO (B)orcdh6ATGMO (C) disrupts apical detachment and EMT. However NCCs C cdh6SplMO NCC Membrane Scale bars: 10 ?m. D cdh6ATGMO NCG Mebrane 00:58:5201:3117 01:5913 (Fig. 3C,D; arrowheads, supplementary material Movie 2). This NCCs were identified as having both green and red fluorescence. We suggests that Cdh6 does not regulate blebbing, and that blebbing alone examined only donor NCCs in areas where the neuroepithelium is not sufficient to generate motility within the neuroepithelium. contained no other donor-derived wild-type cells (n-12 NCCs in Importantly, NCCs showed similar behaviors after injection ofeither four embryos Ofthese, 10 NCCs(83.3%) underwent EMT. Four of morpholino, and the decrease in EMT did not differ between the 10 cells delaminated from the neuroepithelium and initiated morpholinos (P-0.7554 Fisher's exact test), suggesting that the migration during imaging, whereas the remaining six cells were in effects were specific to Cdh6 knockdown. the position of newly delaminated NCCs at imaging onset and To test whether Cdh6 function in EMT is NCC-autonomous, we migrated away from the neuroepithelium. Fig. 4C shows one generated mosaic embryos via blastula stage cell transplantations example of an individual donor NCC undergoing the full process of (Fig. 4A). In donor embryos, all cells were labeled with Texas Red EMT. These data show that individual NCCs with normal Cdh6 dextran and NCCs also expressed GFP [Tg(-4.9sox10:EGFP)]. Host levels can undergo EMT when surrounded by Cdh6-knockdown embryos were injected with Cdh6 morpholinos. Donor cells were cells, indicating that the functions of Cdh6 in detachment and EMT transplanted at 3 hpfand host embryos were imaged at 14 hpf. Donor are NCC-autonomous.

Explanation / Answer

Why? What is the specific question or hypothesis being tested for this figure?

The hypothesis being tested in this case is the the functioning of Cdh6 in the EMT is autonomous or non autonomous.

How? Methods: Describe how the experiment was performed.

In order to do this mosaic embryos had been generated through the transplantation in the blastula stage.

What? Results. Summarize the experimental results, highlighting the relevant controls and main conclusions that can be made only from the data in this figure.

In this experiment, donor cells of NCC had been injected with the cdh6 morpholinos. These had further been identified using red fluorescence and green fluorescence. The donor derived NCC cells showed to undergo EMT. There was initiation of migration at the time of imaging.

So what? Conclusions. Summarize the conclusions from this figure (and only this figure!)

This data implied that the the individual NCC cells having Cdh6 would be undergoing the process of EMT, as thy had been in the presence of Knockdown cells which showed that Cdh6 has its role in detachment of autonomous NCC and EMT.

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