Fig 4. Systemic and cellular phenotypes caused by partial loss of dSTIM in Crz neurons, can be rescued by increasing peptide synthesis or release.

(A) % Pupae upon expression of Insulin receptor (InR) or Ral (Ral^WT) in Crz neurons expressing dSTIM^IR. Genotypes: dSTIM^IR = crz>dSTIM^IR. InR rescue = Crz>InR,dSTIM^IR,dicer2. Ral^WTrescue = Crz> Ral^WT, dSTIM^IR,dicer2. InR control: dicer2;InR/+; dSTIM^IR /+;. Ral^WT control: Ral^WT /+; dSTIM^IR /+. Data represents mean ± SEM. (B) Ral^WTrescue or (C) InR rescue larvae were transferred to normal (Fed) or NR media for 24 hours. Crz immunofluorescence levels in DLP neurons were measured. Number of cells measured shown atop bars. N>7 brains (D) Crz receptor (CrzR) mRNA levels in larval brains expressing various molecules in Crz neurons. InR = Crz>InR. Ral^WT = Crz>Ral^WT. N≥4. (E) % Pupae upon expression of TrpA1 in Crz neurons expressing dSTIM^IR. TrpA1 control: dicer2;TrpA1/+; dSTIM^IR /+; TrpA1 rescue: Crz>TrpA1, dSTIM^IR,dicer2. Larvae were reared at 25°C, at 88h±3h AEL age transferred to NR media and incubated either at 30°C or 22°C for 24 hours, and returned to 25°C thereafter. Data represents mean ± SEM. (F) Crz levels in DLP neurons upon 24 hrs of NR or normal food (Fed) at 30°C for indicated genotypes. N > 5 brains. (G) Crz receptor (CrzR) transcript levels in larval brains expressing various molecules in Crz⁺ neurons, when larvae are reared at 30°C. TrpA1: Crz>TrpA1. N = 6. one-way ANOVA with a post hoc Tukey’s test p<0.05 for (A), (D), (E), (G). Mann-Whitney test for (B), (C). Kruskal-Wallis Test with Dunn’s multi-comparison correction p<0.05 for (F). See also S4 Fig and for source data, S4 Table