N-terminal predicted coiled-coil region of Num1 is required for cortical patch assembly. (A) Widefield single–focal plane images of live cells expressing chromosomally tagged GFP fusion of Num1 constructs. Bars, 1 µm. (B) Western blots of indicated Num1 constructs tagged with GFP. (C) Loss of bright cortical foci in ΔCC1-GFP cells. NUM1-GFP mCherry-TUB1 and ΔCC1-GFP cells, distinguished by mCherry-Tub1 labeling, were imaged in the same field. The percentage of cells exhibiting cortical GFP foci with an intensity >7,700 (arbitrary units) was plotted (n > 228 cells for each strain). Error bars are standard error of proportion. (D) Histograms of fluorescence intensity for individual cortical foci of Num1-GFP, ΔCC1-GFP, ΔTR-GFP, and CC1–303-PH-GFP (n ≥ 173 foci for each construct from a single experiment). denotes mean ± SD. The blue dashed line indicates the mean intensity of Num1-GFP foci. A.U., arbitrary unit.
