Basolateral Endocytic Recycling Requires RAB-10 and AMPH-1 Mediated Recruitment of RAB-5 GAP TBC-2 to Endosomes
Fig 8
Colocalization of recycling cargo on RAB-10 labeled endosomes.
All images are from deconvolved 3D confocal image stacks acquired in intact living animals expressing a GFP-tagged recycling cargo protein, the IL-2 receptor alpha-chain (hTAC-GFP) and tagRFP-RAB-10 in wild-type animals (A-A‴), tbc-2 (tm2241) (B-B‴) and amph-1(tm1060) mutants (C-C‴). (A-A‴) In wild-type animals, hTAC-GFP and tagRFP-RAB-10 colocalizes partially. (B-B‴) In tbc-2 mutants, the degree of colocalization between hTAC-GFP and tagRFP-RAB-10 increased and they mainly overlap on punctate endosomal structures. (C-C‴) In amph-1 mutants, no obvious change in the degree of overlap between hTAC-GFP and tagRFP-RAB-10 was detected in comparison to that in wild-type animals. 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 hTAC-GFP and tagRFP-RAB-10. n = 6. Error bars represent SEM. **P<0.01(student's t test).
