Fig. 3.

Prc::GFP localises to the cardiac ECM but fails to rescue the prc mutations. (A) The cardiac extracellular matrix of a dissected third instar wild-type larva (heart chamber region) stained for the Viking::GFP (anti-GFP, green channel) and endogenously expressed Pericardin protein (anti-Prc, red channel). Viking (Collagen IV) stains all basal laminae whereas Pericardin is restricted to the cardiac matrix. (B) The cardiac extracellular matrix of a dissected third instar wild-type larva (heart chamber region) stained for the Pericardin::GFP fusion protein expressed from the fosmid insertion (anti-GFP, green channel) and endogenously expressed Pericardin protein (anti-Prc, red channel). By staining of the endogenous Prc with anti-Prc, the antibody recognises the Prc::GFP protein as well, which indicates complete overlap. (C) Recombinant prc::GFP^Fosmid, prc^MB03017 animals (C1) were mated with prc^3-21 and prc^3-548. F1 larvae at the third instar larval stage were analysed for partial or complete rescue of the cardiac phenotype (dissociation of pericardial cells as a read out for cardiac ECM disassembly). The analysis was carried out in homozygous animals with the genotype prc::GFP^Fosmid, prc^MB03017/prc::GFP^Fosmid, prc^MB03017 (C1) and animals with the genotypes prc::GFP^Fosmid, prc^MB03017/prc^3-21 (C2) and prc::GFP^Fosmid, prc^MB03017/prc^3-548 (C3). (D) prc^Fosmid was used instead of prc::GFP^Fosmid for the rescue experiment (D1-D3).