Fig. 5. Ectopically expressed mouse PINS can restore apical localization of Insc in mitotic NBs. Anti-PINS antibody can recognize endogenous fly Pins in WT mitotic NBs (A), whereas in pins mutant NBs no signal can be detected (B). Pins is in green and DNA is in cyan. So, although the C-terminal regions of fly Pins and PINS show 33% identity, the anti-PINS raised against the C-terminal region can cross-react with endogenous fly Pins. (C-N) NBs from embryos lacking both the maternal and zygotic components of pins which, in addition, also carry a transgene in which a full-length mouse Pins cDNA is placed under the control of the hsp70 promoter are shown. (C-E) Without heat-shock induction, no PINS protein can be detected in pins mutant background (C) and Insc (red) is cytoplasmic (D); (E) is the merged image of (C) and (D). (F-H) With heat-shock induction, PINS can be detected as an apical crescent in pins mutant mitotic NBs using anti-FLAG staining (F); endogenous Insc can be recruited onto the apical cortex (G); in the merged image (H) it can be seen that Insc and PINS are colocalized as apical crescents in mitotic NBs. (I-K) Heat-shock-induced N-terminal PINS shows cytoplasmic localization (I) and cannot restore Insc apical localization (J) in pins mutant NBs. In the merged image (K) it can be seen that both Insc and N-terminal PINS are localized to the cytoplasm. (L-N) C-terminal PINS is cortically localized in NBs (L), whereas endogenous Insc is still cytoplasmic (M); (N) shows a merged image of (L) and (M).

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