Num1 PH domain is dispensable for cortical patch assembly. (A) Diagram of CC1–303 fusion with GFP or GFP-CAAX. (B) Confocal images of live cells expressing chromosomally tagged GFP constructs expressed by the NUM1 promoter. Bars, 1 µm. (C) The percentage of binucleated cells in cultures grown at 12°C for 16 h (n > 600 cells for each strain). (D) Cortical attachment of mitochondria. Cells expressing Cox4-RFP (red) were fixed and stained for cell periphery with calcofluor (green). Each image is a maximum intensity projection of a 0.8-µm z stack of confocal images. Arrowheads indicate visible association of mitochondria with the cell cortex. Bars, 1 µm. (E) The percentage of cells in which mitochondria were attached to the cell cortex. Cortical attachment of mitochondria was determined from videos of calcofluor-stained cells expressing Cox4-RFP (n ≥ 156 cells for each strain). All error bars are standard error of proportion.
