Basolateral Endocytic Recycling Requires RAB-10 and AMPH-1 Mediated Recruitment of RAB-5 GAP TBC-2 to Endosomes
Fig 6
Loss of TBC-2 or AMPH-1 alters the spatial coordination of RAB-5 and RAB-10.
All images are from deconvolved 3D confocal image stacks acquired in intact living animals expressing GFP- and RFP-tagged proteins specifically in intestinal epithelial cells. (A-A‴) Under wild-type conditions, tagRFP-RAB-5 and GFP-RAB-10 display partial colocalization (~53%) on punctate endosomal structures. (B-B‴) In tbc-2 (tm2241) mutant animals, tagRFP-RAB-5 and GFP-RAB-10 label enlarged pleiomorphic structures and their degree of overlap increased to ~71%. (C-C‴) amph-1 mutants also displayed increased overlap between tagRFP-RAB-5 and GFP-RAB-10 (~76%) with less severe morphological change than that caused by tbc-2 mutants. In each image autofluorescent lysosome-like organelles appears in blue in all three channels, whereas GFP appears only in the green channel and RFP shows up only in the red channel. Signals observed in the green or red channels that do not overlap with signals in the blue channel are considered bona fide GFP or RFP signals respectively. (Scale bar: 10 μm) (D) Pearson's correlation coefficient for colocalization of tagRFP-RAB-5 and GFP-RAB-10. n = 6. Error bars represent SEM. **P<0.01, ***P<0.001(student's t test).
