The SLC-36.1–PPK-3 axis is required for ALR. (A and B) Images of LAAT-1::GFP– and NUC-1::mCh–positive vacuoles in the posterior hypodermis in adult animals of the indicated genotypes without (A) or with starvation (B). Bars, 5 µm. (C) Images of NUC-1::mCh–positive hypodermal vacuoles in N2 and slc-36.1(yq110) adults treated with control (Ctrl) or lgg-1 orlgg-2 RNAi under normal and starvation conditions. Bars, 5 µm. (D) Images of LAAT-1::GFP– and NUC-1::mCh–positive vacuoles in the posterior hypodermis inppk-3(yq24) andatg-2(bp576);ppk-3(yq24) adult animals without or with starvation. Bars, 5 µm. (E) Quantification (mean ± SEM) of the sizes of NUC-1::mCh–positive lysosomes in the posterior hypodermis of adult animals with the indicated genotypes without or with starvation. ***, P < 0.001. (F) Images of NUC-1::mCh–positive hypodermal vacuoles in ppk-3(yq24) adult animals treated with control (Ctrl) or lgg-1 orlgg-2 RNAi under normal and starvation conditions. Bars, 5 µm. (G) Images of NUC-1::mCh–positive hypodermal vacuoles in ppk-3(n2668),slc-36.1(yq110), andslc-36.1(yq110);ppk-3(n2668) adult animals. Bars, 5 µm. (H) Quantification (mean ± SEM) of the sizes of NUC-1::mCh–positive hypodermal vacuoles as shown in G. ***, P < 0.001. (I) Time-lapse chasing of lysosome tubulation on autolysosomes in the hypodermis of N2,slc-36.1(yq110), and ppk-3(n2668) adults. Lysosomal tubulation events are indicated with arrows of different colors. White arrowheads indicate newly formed lysosomes. Bars, 5 µm.