Dorsal pancreatic buds were microdissected from mouse embryos at E12.5 and cultured for up to 3 days at 37°C in 5% CO2. The day of plating is referred to as day 0. For the α6 Integrin inhibition assay, blocking antibodies were added to the culture medium on day 1 at the indicate concentrations. For rescue experiments, both mouse recombinant SLIT1 [500 ng/mL] and SLIT3 (R&D) [1250 ng/mL] were added to the culture medium on day 1. At the end of the experiment, explants cultures were fixed in 4% PFA, stained as whole-mounts as previously described (Petzold and Spagnoli, 2012 (link)).
Slit1
Manufactured by R&D Systems
SLIT1 is a recombinant human protein that functions as a secreted guidance molecule. It plays a role in the development and patterning of various tissues and organs.
Automatically generated - may contain errors
2 protocols using slit1
1
Dorsal Pancreatic Bud Morphogenesis
2
Axon Growth Cone Morphology Assay
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Neurons were treated for 60 min with 100 ng/ml BDNF (PeproTech), 1 μg/μl Ephrin A5 (R&D Systems) or 1 μg/μl Slit-1 (R&D Systems) before fixation. Images were acquired with a Leica TCS SP5 II microscope system and analyses were done with ImageJ (Schindelin et al., 2012 (link)). Growth cone size was measured for determining BDNF effects, whereas repellent cues treated growth cones were categorized into collapsed and non-collapsed according to previous studies (Müller et al., 1990 (link)).
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