FIGURE 7:

rab-10(RNAi) impairs SDPN-1 recruitment to endosomes and segregates RAB-5 from the remaining SDPN-1–labeled structures. All micrographs are from deconvolved 3D confocal image stacks acquired in intact living animals expressing intestinal-specific GFP- and RFP-tagged proteins. Under control conditions (A–A′′′) SDPN-1::GFP and tagRFP::RAB-5 colocalize extensively on endosomes. White arrowheads indicate positive overlap. (C–C′′′) rab-10(RNAi) animals displayed more-diffusive SDPN-1::GFP signal and a decrease in overlap between SDPN-1::GFP and RAB-5::GFP. Gray arrows indicate residual SDPN-1::GFP-labeled structures lacking RAB-5. In each image, autofluorescent lysosome-like organelles appear in in all three channels (including blue), whereas GFP appears only in the green channel and RFP appears only in the red channel. Green and red signals that do not overlap with the blue channel represent pure GFP and RFP signals, respectively. (B) Quantification of SDPN-1::GFP puncta number. (D) Pearson’s r for SDPN-1::GFP and tagRFP::RAB-5. Six animals. Error bars represent SEM: **p < 0.01. Scale bar, 10 μm.