Fig. 3

mir-83 dysregulates autophagy in the intestine through cup-5. a, b GFP::LGG-1 puncta (arrow heads) in the posterior intestine cells (dashed lines) of the WT worms and mir-83(-) mutants at indicated ages. Compared to that of day 1 samples, the exposure of day 5 samples was increased to visualize GFP::LGG-1 puncta. Worms were treated with 5 mM chloroquine (CQ) or a mock control b. n = 3 independent experiments containing at least 22 worms in a and 25 worms in b. c The fluorescent signals of mCherry-GFP::LGG-1 in the posterior intestine cells (dashed lines) of WT worms and mir-83(-) mutants at indicated ages. White arrow heads denote the autolysosome (AL) puncta with only mCherry signals. Yellow arrows denote the autophagosome (AP) puncta with both GFP and mCherry signals. n = 3 independent experiments containing at least 19 worms. d APs (yellow arrows) and ALs (white arrow heads) in the posterior intestine cells (dashed lines) of WT worms and mir-83(-) mutants. Animals were treated with intestine specific RNAi against luc2 or cup-5 from hatching and examined at day 1 of adulthood. n = 3 independent experiments containing at least 16 worms. e The number of SQST-1::GFP puncta (arrow heads) is modestly reduced in the head neurons (dashed lines) of mir-83(-) mutants at the fourth larval stage (L4). n = 3 independent experiments containing at least 58 worms. f The dFP::LGG-1 reporter (mFP/dFP, mFP vs dFP::LGG-1) in neurons responds to the treatment of 5 mM CQ but not mir-83 mutation at day 1 of adulthood. WT samples with control treatment serve as controls for normalization. n = 3 independent experiments. g–h APs (yellow arrows) and ALs (white arrow heads) in the head neurons surrounding the posterior pharynx bulb (dashed lines). Worms of indicated genotypes were examined at day 1 and day 5 of adulthood. A CQ treatment (5 mM) was performed as indicated. n = 3 independent experiments containing at least 15 worms. Scale bars: 10 μm. Statistical significance was calculated by Poisson regression in a–d and g–h, unpaired t-test in e, or one-way ANOVA in f. ns: non-significant, *p < 0.05, ***p < 0.001. Source data are provided as a Source Data file