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. 2007 Feb 12;176(4):483–495. doi: 10.1083/jcb.200608137

Figure 6.

Figure 6.

Localization of anillin relative to prom1 and cadherin in Tis21-GFP–negative versus –positive mitotic NE cells. (A–T) Transverse cryosections of E11.5 forebrain of Tis21-GFP knockin mice (GFP expression; white) double immunostained for anillin (green) and either prom1 (A–J; red) or cadherin (K–T; red) and analyzed by confocal microscopy (C–E, H–J, M–O, and R–T, single optical sections; A, B, F, G, K, L, P, and Q, projection of four consecutive optical sections). Nuclei are stained by DAPI (blue). White asterisks indicate telophase nuclei of daughter cells. Small white bars in M, O, R, and T indicate cadherin hole. (A–E and K–O) Telophase NE cells lacking Tis21-GFP expression and undergoing symmetric division (anillin colocalized with prom1 and cadherin hole upon 90–100% complete ingression of the cleavage furrow). (F–J and P–T) Telophase NE cells showing strong (F) or weak (P) Tis21-GFP expression and undergoing asymmetric division (anillin distinct from prom1 and colocalized with cadherin upon 90–100% complete ingression of the cleavage furrow). (E′, J′, O′, and T′) 3D reconstruction from six to eight 1-μm optical sections showing the prom1–anillin–DAPI merge of the symmetrically dividing Tis21-GFP–negative cell in E (E′), the asymmetrically dividing Tis21-GFP–positive cell in J (J′), the cadherin–anillin–DAPI merge of the symmetrically dividing Tis21-GFP–negative cell in O (O′), and the asymmetrically dividing Tis21-GFP–positive cell in T (T′). White and black arrows in O′ and T′, respectively, indicate the cadherin hole. (U) Quantitation of anillin-stained apical midbodies in symmetrically versus asymmetrically dividing NE cells. Forebrain NE cells of E9.5, E11.5, and E14.5 Tis21-GFP knockin mice were double immunostained for cadherin and anillin and analyzed by confocal microscopy as in K–T. Telophase NE cells showing 90–100% complete ingression of the cleavage furrow (25 out of 54 cells analyzed) were first scored for colocalization of the apical anillin staining with either cadherin-negative (symmetric division) or cadherin-positive (asymmetric division) segments of the cell surface and then for the absence or presence of Tis21-GFP expression. Symmetrically dividing NE cells are expressed as a percentage of symmetrically dividing plus asymmetrically dividing cells; the percentage of symmetrically dividing Tis21-GFP–positive cells is indicated in green. Bar, 5 μm.