CLAMP depletion affects microtubule asymmetry. (A–C) Imaging of microtubules using the EMTB-3xGFP (green) relative to the cell cortex marked with mem-RFP (red) reveals a gap between the microtubules and the cortex at the anterior side of the cell but not the posterior side that is lost in both CLAMP (B) and Vangl2 (C) morphant cells. (D and E) This phenotype was quantified by measuring the mean fluorescent intensity of EMTB-3xGFP within 1 µm of the cell cortex on the posterior side relative to the anterior side (D) in control (n = 33 cells), CLAMP (n = 30 cells), and Vangl2 (n = 48 cells) morphant cells at the level of the basal bodies (E). (F–I) Confocal stacks (∼8 µm deep) of CAMSAP2-GFP (green, F and F′ top and F′ side) reveal a posterior (to the right) bias in MCCs (red; RFP driven by the α-tubulin promoter) that projects across the entire posterior side of the cell (F′ top and side). Mosaic tissue showing a loss of CAMSAP2 (green) asymmetry in CLAMP morphant MCC marked with dextran (blue) compared with a WT MCC (green only; G). Quantification of the CAMSAP2 phenotype measuring the mean fluorescent intensity of CAMSAP2 within the posterior half of the cell relative to the anterior half (H) in control (n = 143 cells), CLAMP (n = 136 cells), and Vangl2 (n = 77 cells) morphant cells (I). Graphs are whisker plots where the error bars represent the range; the line in the box represents the median and the box represents the upper and lower quartile. In all images, posterior is to the right. Bars: 5 µm; (inset) 2.5 µm.