Extended Data Fig. 2: RNA-seq analysis, anorexia in sev > Rasv12 flies, Lgr3Δ50 generation, Lgr3 expression in adult tissues, and eye phenotypes in GMR > ykiSA and the mutants of Dilp8 and Lgr3.

a, RNA-seq analysis showed the increased levels of three tumor-secreted factors (Dilp8, ImpL2, and Upd2) in GMR > yki^SA flies. b, sev > Ras^v12 flies showed anorexia and this phenotype was effectively rescued by Dilp8-RNAi. n = 3 biologically independent experiments; sev > Ras^{v12 *}P = 0.012, sev > Ras^v12 Dilp8-RNAi *P = 0.013, sev > Ras^v12 ImpL2-RNAi *P = 0.019. c, Dilp8 antibody test was performed by western blot and showed almost no expression in the Dilp8^M100727 mutant. d, Dilp8 protein was increased in GMR > yki^SA fly heads and reduced in GMR > yki^SA, Dilp8-RNAi fly heads by western blot. e, The quantification of d using ImageJ program. n = 3 biologically independent experiments; GMR > yki^SA *P = 0.047, GMR > yki^SA Dilp8-RNAi *P = 0.045. f, The Lgr3^Δ50 mutant was obtained using the CRISPR/Cas9-mediated mutagenesis. It consists of 50 base pair deletion in the intron region upstream of target sequences. g, Lgr3 mRNA expression in the Lgr3^Δ50 mutant showed that the Lgr3^Δ50 mutant is a strong hypomorphic allele. n = 3 biologically independent experiments, **P = 0.007. h, GMR > yki^SA,Dilp8^MI00727 and GMR > yki^SA, Lgr3^Δ50 flies respectively blunted or reversed the cancer anorexia phenotype observed in GMR > yki^SA flies. n = 4 biologically independent experiments; GMR > yki^SA *P = 0.010, GMR > yki^SADilp8^M100727 *P = 0.012, GMR > yki^SALgr3^Δ50 *P = 0.027. i, Dilp8 overexpression flies (GMR > Dilp8) showed reduced food intake. n = 6 biologically independent experiments, *P = 0.034. j, In GMR > yki^SA and GMR-Gal4 control flies, the Lgr3 mRNA expression level in the head is very high relative to other tissues. n = 3 biologically independent experiments; head *P = 0.017; thorax **P = 0.006, gut *P = 0.035. k, The eye phenotype in GMR > yki^SA is similar with those of GMR > yki^SA, Dilp8^M100727 and GMR > yki^SA, Lgr3^Δ50. Scale bar is 200μm. l, Lgr3-Gal4 expression marked by GFP. Lgr3 expression was not detectable in sNPF neurons. Scale bar is 100μm. m-n, sNPF (m) (*P = 0.018) and NPF (n) (*P = 0.048) mRNA levels were reduced by RNAi lines with Lgr3-Gal4. n = 3 biologically independent experiments. Data are presented as the mean ± s.e.m. Statistical significance was determined with two-tailed Student’s t-test; *P < 0.05, **P < 0.01. Statistical source data and unprocessed western blots.

Source data