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. 2016 Jan 1;129(1):121–134. doi: 10.1242/jcs.176230

Fig. 1.

Fig. 1.

Pigs is an MT +TIP in Drosophila tissue culture cells and Drosophila tissues in vivo. (A) Schematic of the Pigs protein. CH, Calponin homology domain; Gas2, Gas2 domain; Sx(I/L)P1– Sx(I/L)P3, predicted plus-end MT localisation motifs. (B) GFP–PigsFL (green, B′) localises to the ends of MTs labelled with mCherry–tubulin (red, B″) in S2 cells. Insets show a magnification of the boxed region. (C) GFP–PigsFL (green, C″) partially colocalises with EB1–mRFP (red, C‴) at MT plus-ends in S2 cells. C′–C‴ show a magnification of the boxed region. (D) Frames from a time-lapse movie of a GFP–PigsFL-expressing S2R+ cell shown as a maximum intensity projection of four consecutive frames, pseudocoloured in green (0 s), red (+1 s), blue (+2 s), and then green (+3 s) again. The inset shows a magnified single comet. See Movie 1. (E) Kymograph of GFP–PigsFL tracking an MT plus-end in a S2R+ cell over 16 s. (F) Analysis of comet speeds in S2R+ cells expressing GFP–PigsFL, GFP–PigsCT or GFP–PigsGas2CT. Shown are all data points, the median and the interquartile range. n.s., not significant (Mann–Whitney test). (G) Schematic of Drosophila oogenesis. Pigs was expressed either in squamous follicle cells (green) or the nurse cells of the germline (pink). (H) Still picture from a time-lapse movie of GFP–PigsFL expressed in the squamous follicle cells of a Drosophila ovary in culture at stage 10. Arrows point to plus-end comets marked by GFP–PigsFL. See Movie 2. (I) Kymograph of GFP–Pigs tracking an MT plus-end in the ovary over 15 s. (J) Maximum intensity projection of pseudocoloured frames from a time-lapse movie of ovaries expressing GFP–PigsFL. Green (0 s), red (+1 s), blue (+2 s). Boxes indicate single comets that are shown magnified to the right. (K) Still from a time-lapse movie of Drosophila ovary squamous follicle cells at stage 10 expressing GFP–PigsFL (green, K′) at the end of MTs labelled by mCherry–CLIP170 (mCh-CLIP; red, K″). Arrow points to a comet, arrowhead points to Pigs colocalisation with lattice MTs. K′–K″ show a magnification of the boxed region. (L) Kymograph of GFP–PigsFL (L′, green) and mCherry–CLIP170 (L″, red) tracking an MT plus-end in the ovary squamous follicle cells over 12 s. (M) Analysis of comet speeds in vivo in squamous follicle cells expressing EB1–GFP, mCh-CLIP, GFP–PigsFL or GFP–PigsCT. Shown are all data points, the median and the interquartile range. Significance was tested using Mann–Whitney test; n.s, not significant. Scale bars: 5 µm (B–D, H, J, K); 1 µm (E, I , L).