Extended Data Fig. 6: Conservation of INSL3/Lgr8 in mammals, and cachexia-anorexia phenotype of C26 or LLC implanted mice.
a. The phylogenetic comparative analysis of Drosophila Lgr3 and mammalian Lgr family shows that Lgr7/8 is a homologue of Lgr3. b, The phylogenetic comparative analysis of fly Dilp8 and mammalian INSL3 shows that they are classified as close peptides. c, Lgr8 protein is strongly expressed in the hypothalamic region and hippocampus of the mouse brain. d, In the RNA FISH, Lgr8 mRNA is strongly expressed in the hypothalamic region and hippocampus of the mouse brain. e, Lgr8 is expressed in the mouse hypothalamic N3 cells, but not in N39 cells. f-g, Body weight was changed in C26 tumor implanted mice (f) not in LLC tumor implanted mice (g). n = 7 mice for both groups. f, 2w + 2day *P = 0.027, 2w + 4day ***P = 0.0004, 2w + 6day **P = 0.0034. h, Tumor volume and weight changes were measured in C26 and LLC implanted mice. n = 6 mice for both groups. i-j, Muscle and fat weight of C26 tumor implanted mice at day 12 (i) and day 21 (j). Tibialis anterior (TA), gastrocnemius (GCM), inguinal white adipose tissue (iWAT), and epididymal white adipose tissue (eWAT). Control n = 3 mice, C26 day 12 n = 5 mice, C26 day 21 n = 4 mice. i, iWAT *P = 0.024, eWAT *P = 0.094. j, GCM *P = 0.038, iWAT **P = 0.001, eWAT *P = 0.013. k-l, Nucb2 (k) and Npy (l) mRNA levels were not changed in LLC implanted mice. n = 5 mice for both groups. m, INSL3 i.p. injected wild-type mice didn’t show reduction of food intake. n = 7 mice for both groups. n-o, Nucb2 (n) and Npy (o) mRNA levels were not changed in INSL3 i.p. injected wild-type mice hypothalamus. n = 3 mice for both groups. 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, ***P < 0.001. Statistical source data and unprocessed western blots.
