Fig. 4

Quantification of dorsal versus ventral cilia emergence. pH Smo expressing cells (green) were labeled with the anti-GFP antibody (red) and Hoechst dye (blue) in fixed, unpermeabilized cells. The cells were then imaged either closer to the coverslip via TIRFM or slightly deeper towards the middle of the cell by HILO within the same field of view. a Two cilia in adjacent cells can be seen: one outside near the bottom of the cell seen by TIRFM (yellow zoom-in box, marked by *V for ventrally emerged), the other inside within the middle of the cell only visible by HILO (white zoom-in box, marked by M for middle). b A dorsal, partially emerged cilia (marked by *D) not visible by TIRFM, but imaged via HILO. Asterisk marks emerged cilia. c Left schematic of microscopy approaches used and the relative cell depth visualization. Right schematic of Dorsal (D), Middle (M) and Ventral (V) cilia emergence classification. Middle cilia are classified as being roughly 0.5 μm away from the membrane. d Quantification of the frequencies of cilia emergence relative to cell position from SDCM data