Skip to main content
. 2015 Jan 23;4(2):233–243. doi: 10.1242/bio.201410827

Fig. 3. NetB and Fra endocytosis and colocalisation.

Fig. 3.

Stage 13 embryos immunostained for Fas3 (blue A–F), NetB (red A–C,I; blue L; grey A′,B′,C″,J′,K′,L″), Fra (red, D–H; grey D′,E′,F″–H″), GFP (green, A–H,L; grey C′,F′,G′,H′,L′) and HA (green I; grey K,L″′). (A) In embryos expressing YFP-Rab5 (green), NetB (red) is enriched in the basal regions of the midgut (arrow) (n = 5). (B) This line is greatly reduced in embryos expressing dominant-negative YFP-rab5S43N (B′, arrow) (n = 8). (C) NetB colocalises with YFP-Rab5 extensively in basal regions of the cell, and in some internal puncta (yellow arrowheads) (n = 5). (Note: some puncta express only NetB/Fra (red arrowheads), while others express only Rabs (green arrowheads) indicating that colocalisation in C, F and G is not due to “bleedthrough” between channels). (D) In embryos expressing YFP-Rab5 (green), Fra (red) is enriched in the basal regions of the midgut and in many internal puncta (arrowheads) (n = 8). (E) In embryos expressing dominant-negative YFP-rab5S43N internal puncta are absent and stronger lateral membrane staining is seen (E′, arrow) (n = 5). (F) Fra colocalises with Rab5 in many internal puncta (yellow arrowheads) (n = 8). (G) Fra colocalises with Rab7-GFP expressing late endosomes (n = 5). (H) No colocalisation was seen with the recycling endosomal marker Rab11-GFP (n = 7). (I–K) Co-localisation between NetB and Fra. A stage 13 embryo 48Y-GAL4/+;UAS-fra-HA/+ embryo, stained with HA (green) to show Fra expression and NetB (red) (n = 4). (I) Co-localisation between NetB and Fra was seen predominately on the basal side of the midgut cells. (J–J′). Zoomed image of the region indicated by the right box in I. Arrowheads indicate co-localisation between NetB and Fra on the basal side of the midgut. (K–K′) Zoomed image of the left box in I. NetB and Fra-HA colocalise in some internal punctae. (L) Stage 13 embryo co-expressing Rab5-GFP and Fra-HA shows colocalisation of NetB, Rab5 and Fra in some internal puncta (arrowheads) (n = 5). Scale bars, 20 µm (A,B,D,E); 10 µm (C,F,G,H,J,K,L).