CLAMP is required cell autonomously for intercalation. (A) Mosaic embryos coinjected with CLAMP-MO and αtub-GFP (arrows) stained with anti-acetylated tubulin and phalloidin. (B) Mosaic embryos coinjected with CLAMP-MO and membrane-RFP stained with anti–β-tubulin and phalloidin (wild-type MCCs still intercalate, arrows). (C) Mosaic embryo coinjected with CLAMP-MO and membrane-RFP (blue), showing MCC intercalation defects (arrows). (D) Quantification of intercalation in MCCs injected with control MO, CLAMP MO, or CLAMP MO rescued with xCLAMP or hCLAMP. (E and F) Representative image (E) and quantification (F) of intercalation defect in ICs (green, AE1 staining) containing CLAMP MO (arrowheads) versus wild-type (arrow, lack of red). (G and H) Image (G) and quantification (H) of centriole position in CLAMP morphant MCCs (the arrow highlights the centriole cluster). (I and J) Representative image (I) and quantification (J) of mosaic embryos showing the loss of CLAMP staining in DN-Par3–expressing MCCs (arrowheads) compared with wild-type (arrow). Quantification in H is based on at least 10 cells, each from a total of at least five embryos from at least two independent experiments. Quantifications of intercalation defects in CLAMP morphants in D and F are significantly different from controls, as are morphant embryos from rescue (P < 0.0001, see Table S1). A χ2 test shows no statistical significant difference in H (P = 0.236, see Table S1). Quantification of reduction in apical CLAMP levels in DN-Par–expressing cells is significantly different from controls (P < 0.01, see Table S1). See Fig. S3. Error bars represent standard deviations. Bars, 5 µm.
