Figure 6.
RAB-11 recruits RABI-8 to vesicular endosomal compartments and regulates unconventional apical protein transport. (A) Confocal images of intestinal cells expressing GFP-tagged RABI-8. Compared with wild-type animals, rab-11 knockdown significantly reduced the number and intensity of RABI-8::GFP puncta, while mutation in rab-8 had no obvious effect. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a one-way ANOVA followed by a post-hoc test (Dunn’s Multiple Comparison Test) for multiple comparisons. ***P < 0.001; ns, no significance. Data distribution was assumed to be normal but this was not formally tested. Scale bars: 10 μm. (B) RABI-8 precipitated GFP::RAB-11 in a representative Co-IP assay. (C) Confocal images of intestinal cells expressing GFP-tagged PGP-1. Compared with wild-type animals, GFP::PGP-1 accumulated in intracellular structures in rab-11(RNAi) knockdown animals. For quantification, the signals from the apical membrane were avoided by manual ROI selection. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a two-tailed, unpaired Student’s t test. ***P < 0.001. Data distribution was assumed to be normal but this was not formally tested. About one cell length of the intestine is shown in each panel. Scale bars: 10 μm. Yellow asterisks indicate intestinal lumen. Dashed lines indicate the outline of the intestine. (D) Confocal images of wild-type and rab-11(RNAi) knockdown animals after a 48-h incubation with fluorescent dye rho123. Accumulation of rho123 was about 10-fold higher in rab-11(RNAi) than in wild-type animals. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a two-tailed, unpaired Student’s t test. ***P < 0.001. Data distribution was assumed to be normal but this was not formally tested. Scale bars: 20 μm. (E) Compared with wild-type animals, rab-11 knockdown reduced the survival rate after 24 h of P. aeruginosa exposure. PGP-1 overexpression (OE) improved the survival rate of rab-11(RNAi) animals. (F) Compared with wild-type animals, rab-11 knockdown reduced the survival rate upon colchicine (2 mM) treatment. PGP-1 overexpression improved the survival rate of rab-11(RNAi) animals. Statistical significance was determined using a one-way ANOVA followed by a post-hoc test (Dunn’s Multiple Comparison Test) for multiple comparisons. **P < 0.01; ***P < 0.001; ns, no significance. Data distribution was assumed to be normal but this was not formally tested. Source data are available for this figure: SourceData F6.

RAB-11 recruits RABI-8 to vesicular endosomal compartments and regulates unconventional apical protein transport. (A) Confocal images of intestinal cells expressing GFP-tagged RABI-8. Compared with wild-type animals, rab-11 knockdown significantly reduced the number and intensity of RABI-8::GFP puncta, while mutation in rab-8 had no obvious effect. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a one-way ANOVA followed by a post-hoc test (Dunn’s Multiple Comparison Test) for multiple comparisons. ***P < 0.001; ns, no significance. Data distribution was assumed to be normal but this was not formally tested. Scale bars: 10 μm. (B) RABI-8 precipitated GFP::RAB-11 in a representative Co-IP assay. (C) Confocal images of intestinal cells expressing GFP-tagged PGP-1. Compared with wild-type animals, GFP::PGP-1 accumulated in intracellular structures in rab-11(RNAi) knockdown animals. For quantification, the signals from the apical membrane were avoided by manual ROI selection. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a two-tailed, unpaired Student’s t test. ***P < 0.001. Data distribution was assumed to be normal but this was not formally tested. About one cell length of the intestine is shown in each panel. Scale bars: 10 μm. Yellow asterisks indicate intestinal lumen. Dashed lines indicate the outline of the intestine. (D) Confocal images of wild-type and rab-11(RNAi) knockdown animals after a 48-h incubation with fluorescent dye rho123. Accumulation of rho123 was about 10-fold higher in rab-11(RNAi) than in wild-type animals. Data are shown as mean ± SD (n = 18 each, six animals of each genotype sampled in three different unit regions of each intestine defined by a 100 × 100 [pixel2] box positioned at random). Statistical significance was determined using a two-tailed, unpaired Student’s t test. ***P < 0.001. Data distribution was assumed to be normal but this was not formally tested. Scale bars: 20 μm. (E) Compared with wild-type animals, rab-11 knockdown reduced the survival rate after 24 h of P. aeruginosa exposure. PGP-1 overexpression (OE) improved the survival rate of rab-11(RNAi) animals. (F) Compared with wild-type animals, rab-11 knockdown reduced the survival rate upon colchicine (2 mM) treatment. PGP-1 overexpression improved the survival rate of rab-11(RNAi) animals. Statistical significance was determined using a one-way ANOVA followed by a post-hoc test (Dunn’s Multiple Comparison Test) for multiple comparisons. **P < 0.01; ***P < 0.001; ns, no significance. Data distribution was assumed to be normal but this was not formally tested. Source data are available for this figure: SourceData F6.

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