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. 2018 Jun 4;7:e33286. doi: 10.7554/eLife.33286

Figure 6. Peroxisomal co-localization of ACS-7, ACOX-1.1, and ACOX-3.

(A) Co-injection of plasmids containing Pacox-1.1::mcherry::acox-1.1 and Pacs-7::gfp::acs-7 into wild-type worms shows that ACOX-1.1 and ACS-7 co-localize in a punctate pattern in the intestine. (B) Co-injection of plasmids containing Pacox-1.1::mcherry::acox-1.1 and Pacox-3::gfp::acox-3 into wild-type worms shows that ACOX-1.1 and ACOX-3 co-localize in a punctate pattern in the intestine. (C) RNAi using the control plasmid L4440 in Pacox-1.1::mcherry::acox-1.1;Pacs-7::gfp::acs-7 worms gives a punctate pattern of expression in the intestine. (D) RNAi against prx-13 in Pacox-1.1::mcherry::acox-1.1; Pacs-7::gfp::acs-7 worms gives a diffuse pattern of expression in the intestine. In (A–D), L4 to young adult stage worms were imaged. Scale bar = 20 μm.

Figure 6.

Figure 6—figure supplement 1. ACS-7 does not localize primarily to the lysosome.

Figure 6—figure supplement 1.

(A) Staining of Pacs-7::gfp::acs-7 L4 worms with LysoTracker Red dye. (B) Staining of Pacs-7::gfp::acs-7 adult worms with LysoTracker Red dye. (C,D) Staining of Pacs-7::gfp::acs-7 adult worms with LysoTracker Red dye, before (C) and after (D) bleaching the GFP signal under the fluorescent microscope. Bleaching in (C) selectively eliminates peroxisomal GFP while making lysosomal autofluoresence more prominent. (E) Staining of wild-type (N2) worms with LysoTracker Red dye. In (A–E), peroxisomes are indicated with arrows and lysosomes are indicated with triangles. In (A), L3 worms are imaged, and in (B–E), L4 to adult worms are imaged. Scale bar = 20 μm.