Fig. 4. Sugar transporter 2 in the EECs regulates glucose-stimulated NPF secretion in mated females.
a,b, Feeding time measured using FLIC. a, n = 16 EEC>, n = 12 EEC>sut2i. b, n = 7 NPFgut>, n = 10 NPFgut>NPFiTRiP, n = 11 NPFgut>sut2i, n = 17 NPFgut>Mondoi. c, Sugar consumption measured by CAFÉ assay; n = 14 NPFgut>, n = 15 NPFgut>sut2i, n = 15 NPFgut>Mondoi. d, Consumption of sucrose or yeast measured by dye assay. Sucrose n = 22 NPFgut>, n = 16 NPFgut>sut2i. Yeast n = 10 NPFgut>, n = 15 NPFgut>sut2i. e, Consumption preference measured by two-choice dye-consumption assay. Virgins, n = 25 NPFgut>, n = 17 NPFgut>sut2i; mated n = 24 NPFgut>, n = 19 NPFgut>sut2i. f, Midgut NPF staining with sut2 knockdown; n = 571 cells from six guts for NPFgut>, n = 625 cells (five guts) for NPFgut>sut2i. g,h, Representative images of midgut NPF-cell activity (g), quantified in h; left shows n = 1,167 cells from eight fed guts, n = 1,394 cells from nine starved (stv) guts; right shows n = 1,209 cells from eight fed guts, n = 1,405 cells from nine starved guts. i, Midgut NPF expression; n = 6 for stv, 0.5 h, 1 h, 4 h and n = 5 for 2 h, 6 h, each samples of five guts. j, Representative images of NPF staining and cell activity in 24-h starved, 2-h sugar re-fed and 6-h sugar re-fed mated females. All measured cells are marked with tdTomato. k, Quantification of j. Eight guts per condition. Left, n = 1,297, 1,038 and 1,216 cells from starved, 2-h and 6-h re-fed animals. Right, n = 1,141, 954 and 1,151 cells from starved, 2-h re-fed and 6-h re-fed animals. l, Midgut NPF expression in mated females after re-feeding following 24-hour starvation (Stv, no re-feeding); n = 6 replicates of six midguts except n = 5 replicates for 2 and 6 h. m, NPF staining in mated females’ midguts after 15 hours’ knockdown of Mondo (29 °C inactivation of GAL80TS) and following subsequent 6-h derepression of Mondo (18 °C). 15 h at 29 °C: n = 1,595 cells/12 guts for EEC>, n = 1,840 cells/14 guts for EEC>Mondoi; 6 h at 18 °C: n = 1,948 cells/17 guts for EEC>, n = 2,194 cells/18 guts for EEC>Mondoi. All animals were mated females, except in e. Bars represent mean ± s.e.m. Box plots indicate minimum, 25th percentile, median, 75th percentile and maximum values. NS, not significant. a,d (left), Two-tailed unpaired Student’s t-test. b,e,k, One-way Kruskal–Wallis ANOVA with Dunn’s multiple-comparisons test. c,l, One-way ANOVA with Dunnett’s multiple-comparisons test. d (right), f,h,i,m, Two-tailed unpaired Mann–Whitney U-test. Scale bars, 50 μm.