Figure S5.
Kif6 deficiency disrupts rotational polarity but not the translational polarity of BBs in the ependyma. Related to Fig. 7. Se, Severe hydrocephalus; Mi, Mild hydrocephalus. (A and B) BBs in Kif6−/− but not in Kif9−/− ependymal cells displayed impaired rotational polarity. Ependymal tissues from the indicated littermates were co-immunostained for Cep164 to visualize BBs and Centriolin to visualize BFs. Framed regions (A) were magnified to show BF orientations (arrows). The extent of the rotational polarity for a BB patch (BBs in a cell) was calculated as the mean vector length of all the BBs with clear BFs by considering each arrow marking their polarities as a unit vector. Quantification results from three mice per genotype are presented as mean ± SD plus sample dots and subjected to unpaired two-tailed student’s t test. Mice used were two 6-month-old (6 M) and one 7-month-old mice for Kif9−/− and three of their wild-type littermates; three P35 mice with severe hydrocephalus for Kif6−/− and three of their wild-type littermates. 20 ependymal cells (BB patches) were scored for each mouse. (C) Representative 3D-SIM images of ependymal tissues from the indicated mice. BB patches magnified from the framed regions are presented in A. Arrows indicate directions of the rotational polarity of BB patches, based on major polarities of the BBs. Note that the arrows in each micrograph point to similar directions. No arrows are drawn for the Kif6−/− BB patches due to their lack of rotational polarity (B). (D)Kif6 or Kif9-deficiency did not alter BB number in ependymal cells. 50 cells were measured per mouse. (E–H)Kif6 deficiency did not impair the translational polarity of BBs. Ependymal tissues from Kif6−/− mice with mild hydrocephalus (two P62 and one P68) and severe hydrocephalus (P34, P38, and P43) or wild-type mice (P62, P38, and P43) were immunostained for ZO1 and Cep164 to visualize cell borders (tight junctions) and BBs, respectively (E). BB patch displacement relative to the cellular geographic center (G) and relative BB patch area (H) were quantified as illustrated in (F). 50 ependymal cells were scored for each mouse.

Kif6 deficiency disrupts rotational polarity but not the translational polarity of BBs in the ependyma. Related to Fig. 7. Se, Severe hydrocephalus; Mi, Mild hydrocephalus. (A and B) BBs in Kif6−/− but not in Kif9−/− ependymal cells displayed impaired rotational polarity. Ependymal tissues from the indicated littermates were co-immunostained for Cep164 to visualize BBs and Centriolin to visualize BFs. Framed regions (A) were magnified to show BF orientations (arrows). The extent of the rotational polarity for a BB patch (BBs in a cell) was calculated as the mean vector length of all the BBs with clear BFs by considering each arrow marking their polarities as a unit vector. Quantification results from three mice per genotype are presented as mean ± SD plus sample dots and subjected to unpaired two-tailed student’s t test. Mice used were two 6-month-old (6 M) and one 7-month-old mice for Kif9−/− and three of their wild-type littermates; three P35 mice with severe hydrocephalus for Kif6−/− and three of their wild-type littermates. 20 ependymal cells (BB patches) were scored for each mouse. (C) Representative 3D-SIM images of ependymal tissues from the indicated mice. BB patches magnified from the framed regions are presented in A. Arrows indicate directions of the rotational polarity of BB patches, based on major polarities of the BBs. Note that the arrows in each micrograph point to similar directions. No arrows are drawn for the Kif6−/− BB patches due to their lack of rotational polarity (B). (D)Kif6 or Kif9-deficiency did not alter BB number in ependymal cells. 50 cells were measured per mouse. (E–H)Kif6 deficiency did not impair the translational polarity of BBs. Ependymal tissues from Kif6−/− mice with mild hydrocephalus (two P62 and one P68) and severe hydrocephalus (P34, P38, and P43) or wild-type mice (P62, P38, and P43) were immunostained for ZO1 and Cep164 to visualize cell borders (tight junctions) and BBs, respectively (E). BB patch displacement relative to the cellular geographic center (G) and relative BB patch area (H) were quantified as illustrated in (F). 50 ependymal cells were scored for each mouse.

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