Temporal control of axonal floor plate crossing through a combination of incoherent feedforward and feedback loops of gene regulatory network regulating Robo3 expression

Description

Excell files that includes data about number of cell, expression levels and axonal projections.

Steps to reproduce

Quantification of commissural projection Images of spinal cord cross sections were analyzed using ImageJ (National Institutes of Health, USA) tools. Specifically, the number of pixels at the white matter on the ipsilateral and the contralateral spinal cord (without the floor plate) was scored. % comm = #commissural pixels / (#commissural pixels + ipsilateral pixles). Analysis of Barhl2 expression in Lhx2 Lhx9 double null mice The Lhx2;Lhx9 mouse embryo tissue was generated in Professor Jane Dodds lab, Columbia University under approval by the IACUC and gifted for reuse. 3 E11.5 Lhx2+/-;Lhx9+/- and 9 control embryos were analyzed. Controls consisted of embryos which were either wildtype, single or double heterozygote for the Lhx2 and Lhx9 genes. Math1LacZ transgene was present in some embryos. Transverse spinal cord sections were labeled by immunohistochemistry with Barhl2 and DAPI and images acquired at subthreshold intensity. The raw images were analyzed in Cell Profiler (www.cellprofiler.org) where ‘objects’ (Barhl2+ cells) were delineated and the pixel count in each Barhl2+ cell quantified. This resulted in a pixel distribution of Barhl2+ cells in the spinal cord of control and double mutant embryos. As three different litters cohorts were analyzed, the data was normalized to control values within the same litter in order that data acquired form separate litter cohorts could be accurately compared. The data was statistically analyzed using Prism 9. Raw data pixel counts are available on request.

Institutions

• Hebrew University of Jerusalem

  Jerusalem, Jerusalem

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