Figure 6

Lva-marked Golgi vesicles display increased size in Pld^nullembryos and plasma membrane insertion of Neurotactin (Nrt) is defective, but Rab11 localization is unaltered. Cellularizing embryos were immunostained with affinity-purified antisera to visualize trafficking proteins and subcellular compartments. A, B. Anti-Lva antisera to visualize Golgi vesicles A. Two y¹w^67c23control embryos, representative of 82 examined, illustrate Lva-stained vesicles present both apically and basally. B. Two Pld^nullembryos, representative of 75 examined, reveal an increased intensity of Lva staining and increased size of Lva-marked Golgi vesicles. Images were taken at or near the embryonic equator. The differences in Lva-staining intensity were reproducible across multiple experiments and batches of embryos. Aggregate size distributions were quantified on 12 y¹w^67c23control and 12 Pld^nullembryos using NIH Image J software. C, D. Cellularizing embryos were heat-fixed and immunostained with mouse monoclonal anti-Nrt. C. In representative y¹w^67c23control embryos (two are shown), Nrt accumulates at the plasma membrane and in small vesicles. D. In representative Pld^nullembryos, in contrast, neurotactin accumulates subapically and apicolaterally. In total, 36 embryos from multiple experiments were examined with similar results. E, F. Cellularizing embryos were heat-fixed and immunostained with rat polyclonal anti-Rab11.