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Expression of collagen IV and Pxdn during development in zebrafish and morpholino (MO) knockdown of peroxidasin in zebrafish embryos. (A) Pxdn and collagen IV expression during zebrafish embryonic development. Real-time qPCR studies were conducted to examine expression levels of Pxdn, collagen4α1, and collagen4α2. *Student t test P value < 0.03 compared with expression at 1,000 cells. Error bars = SEM. Blue, pxdn; red, col4a; black, col4a2. (B) Control and (C) Pxdn MO groups. MO-injected embryos displayed (D) general severe defects that include cardiac edema, smaller eyes, and gross trunk patterning defects (4/45), (E) partial curved trunk (21/45), or (F) normal development (20/45). (G) SDS/PAGE analysis of Pxdn MO embryonic phenotypes at 24 hpf by Western blot. Collagenase digests were normalized for total protein load by protein stain with SYPRO-Ruby (Fig. S8).
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