XB-IMG-151250
Xenbase Image ID: 151250
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Figure 3. Presence of iduronic acid in chondroitin/dermatan sulfate
proteoglycans of early embryos.
(A) At stage 22, IdoA is present in high molecular weight CS/DS-PGs, as
demonstrated by the size-fractionation of [35S]-PGs. The HS and CS/DS degradation products are produced by nitrous acid and Chase ABC treatment, respectively. CS/DS-PGs represent 72% of the high molecular weight-PGs (fractions 12-17).
(B) SDS-PAGE analysis of [35S]-labeled CS/DS-PGs. The same samples analyzed by gel filtration (A) were separated using a 4-10% gradient SDS-PAGE following nitrous acid and alternatively Chase ABC or Chase B treatments. The fluography indicates CS/DS-PGs (brackets) with an apparent molecular weight of 200-300 kDa (Bgn) and c. 1,000 kDa (Vcan). The percentages of radioactivity in the framed areas are
indicated below each lane.
(C,C’) Whole-mount in situ hybridization of Bgn at stage 26. Embryo is shown in the lateral view (C) and transversally sectioned (C’). Arrowheads indicate migrating trunk neural crest cells.
(D,E) Chase B treatment degrades CS/DS chains in high molecular weight-PGs in control-MO- (D) but not Dse-MO-injected embryos (E).
(F) qPCR analysis of CNC explants at stage 18. Dse-MO does not differentially affect the mRNA levels of Dse and Dsel compared with Dse-5MM-MO. Mean ±SDEV from triplicates. Number of biological replicates n=4. Bgn, biglycan; Chase, chondroitinase; Vcan, versican. Image published in: Gouignard N et al. (2016) © 2016. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license
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