Figure 7—figure supplement 6.

Supplementary data on the ArhGEF11/PDZ-RhoGEF CRISPR C-ter deletion phenotype.

(Kdrl:eGFP-Jam3b; kdrl:ArhGEF11^{CRISPR-Cterdel+/+}) homozygous ArhGEF11 C-ter deletion mutants (CRISPR ArhGEF11: panels B, D, F) and control siblings (Control: panels A, C, E) were imaged using spinning disk confocal microscopy (48-55hpf time-window). Note that the genetic background of the CRISPR fish line is (Kdrl:GAL4; UAS:RFP) thus allowing the red cytosolic staining of aortic and hemogenic cells. Aorta segments (330 µm each) were imaged in the trunk region. The 2D-cartographies with delimited cellular contours in panels (A and B) are presented Figure 7B with the corresponding un-modified 2D-cartographies shown on top for unmasking of contours. Panels (A, C) and (D, F) illustrate 2 different segments from the same embryo. (a–f) Maximum z-projections of merged nls-mkate2 and eGFP-Jam3b signals for control (a, c, e) and for the CRISPR mutant (b, d, f) conditions. For panels (a and b), maximum z-projections of the eGFP-Jam2a signal only are also shown. (a’-f’) Single z-plane images of merged nls-mkate2 and eGFP-Jam3b signals for control (a’, c’, e’) and for ArhGEF11 exon 38 splicing morpholino (b’, d’, f’) conditions. For b’ and d’, the image is a composition of 2 different z-planes from the same field (the boundaries are marked with white ticks). In right margins, magenta and green arrowheads designate the aortic floor and roof, respectively. (a’’-f’’) 2D-cartographies (bottom, with delineated cellular contours) obtained from eGFP-Jam3b signals for control (a’’, c’’, e’’) and for the CRIPR mutant (b’’, d’’, f’’) conditions, respectively. Cell contours are delineated either in blue (endothelial cells), yellow (hemogenic cells), red (morphologically characterized EHT cells, red arrows), and small cells delineated by cyan boxes (morphologically uncharacterized EHT cells). Cellular contours have been semi-automatically segmented along the cellular interfaces labeled with eGFP-Jam3b (see Materials and methods). White and black arrows designate hemogenic cells with their nucleus visible on the z-section. Analyses were performed on 2 x wild-type siblings for controls and 2 x mutant embryos whose mutation was confirmed by DNA sequencing. Scale bars = 20 µm.