Figure 7.

LLC-PK1 cells have a non-polarized distribution of EGFR and IQGAP1, and they consequently suffer spindle misorientation.

A, B LLC-PK1 cysts were grown for 72 h, fixed and then stained for (A) EGFR (green), actin (red) and β-catenin (blue), or (B) IQGAP1 (green), actin (red) and β-catenin (blue). The apical localization of endogenous EGFR or IQGAP1 is indicated (arrowheads).

C Percentage of cysts forming a single lumen: MCDK, 82.96 ± 3.17%; LLC-PK1, 51.86 ± 5.94% (n = 3, >100 cysts per experiment; values represent the mean ± s.d.).

D LLC-PK1 cysts were stained for acetylated tubulin (green), actin (red) and β-catenin (blue). The yellow line represents mitotic spindle angle.

E Mean spindle angles: MDCK = 74.73 ± 3.52°, LLC-PK1 = 45.37 ± 5.07° (n = 3, 30 cysts per experiment; values represent the mean ± s.e.m.).

F, G LLC-PK1 AP1B cysts were grown for 72 h, fixed and then stained for (F)EGFR (green), actin (red) and β-catenin (blue), or (G) IQGAP1 (green), actin (red) and β-catenin (blue). The basolateral localization of endogenous EGFR or IQGAP1 is indicated (arrows).

H The percentage of LLC-PK1-AP1B cysts forming a single lumen was quantified: LLC-PK1-AP1B, 65.57 ± 5.23% (n = 3, >80 cysts per experiment; values represent the mean ± s.d.).

I LLC-PK1-AP1B cysts were stained for acetylated tubulin (green), actin (red) and β-catenin (blue). The yellow line represents mitotic spindle angle.

J Mean spindle angles: LLC-PK1 = 45.29 ± 5.09°, LLC-PK1 AP1B = 61.80 ± 5.30° (n = 3, 30 cysts per experiment; values represent the mean ± s.e.m.).

Data information: Error bars represent the s.d. or s.e.m: *P < 0.05; **P < 0.01. Scale bars, 5 μm.