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Fig. 4. Collagen IV associates with basal laminae in SPARC mutants that express transgenic SPARC. Embryos transgenically expressing SPARC were immunostained with anti-collagen IV (ColIV, red) and anti-SPARC (dSPARC, green) antibodies. (A) ES16 embryo expressing the SPARC transgene in a SPARC mutant background in haemocytes (UAS-SPARC; gcm-GAL4) shows SPARC colocalization with collagen IV in basal laminae (arrow, yellow). No morphological defects are observed. Endogenous SPARC expression and colocalization with collagen IV is observed in the fat body (arrowhead). (B) ES16 rescued embryo (UAS-SPARC/+ or y; gcm-GAL4/+; H2Av-GFP, Df(3R)nm136/H2Av-GFP, Df(3R)nm136) immunostained for SPARC (green). (C) The same embryo as in B immunostained for collagen IV (red). GFP expression is not shown. (D) The merged image (yellow) shows their colocalization in haemocytes (arrowhead) and basal laminae (arrows) around the brain, VNC, midgut chambers and along the VNC channels. (E) Lateral view of an ES16 SPARC-mutant embryo that expresses transgenic SPARC in haemocytes (UAS-SPARC/+ or y; SrpHemo-GAL4 UAS-GFP/+; H2Av-GFP, Df(3R)nm136/H2Av-GFP, Df(3R)nm136) immunostained for collagen IV (red) and SPARC (green). Intense coimmunostaining is observed in haemocytes (arrowheads) and basal laminae (arrows). (F) ES17 embryo with the same genotype as in E. Collagen IV and SPARC colocalize in basal laminae. (G) UAS-SPARC/+ or y; gcm-GAL4/+; H2Av-GFP, Df(3R)nm136/H2Av-GFP, Df(3R)nm136 embryos immunostained for laminin (red). Laminin distribution is continuous around the VNC. (H) Lateral view of an ES17 SPARC-mutant embryo expressing SPARC (blue) in all neuroblast- and glia-derived cells and sensory organ precursor cells (UAS-SPARC/+ or y; sca-GAL4/+; H2Av-GFP, Df(3R)nm136/H2Av-GFP, Df(3R)nm136). Motorneurons are disorganized and the VNC twists out of focus. All images are single confocal sections.
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