Figure 5. Role for Rh6 for suppression of sucrose appeal, and expression of Rh6 in a subset of bitter GRNs.
(A) PER assays to screen for contributions of candidate signaling proteins to cool suppression of sugar attraction. Flies were presented with 0.5 M sucrose at either 23° or 17°C. Shown are the 3rd PER offerings. See Figures S6A and S6B for the results of the 1st and 2nd offerings. n=5 trials. Each trial includes ≥8 flies. Non-parametric Mann-Whitney test. *P≤0.05. Error bars represent means ±SEMs. See Figure S7A—S7D.
(B—I) Staining control labella with anti-Rh6, which labels bitter GRNs. UAS-tdTomato was expressed in bitter GRNs under the control of the Gr66a-Gal4.
(B—E) Z stacks focusing on S-type sensilla close to the inner surface of the labellum.
(B) Anti-Rh6 staining (green). Scale bar represents 20 μm.
(C) tdTomato fluorescence (red).
(D) Merge of B and C.
(E) tdTomato fluorescence superimposed on DIC image of the labellum. S-type sensilla are labelled. S6 is slightly longer than other S-type sensilla. The S-type nomenclature used is as described [56].
(F—I) Z stacks focusing on I-type sensilla close to the outer surface of the labellum.
(F) Anti-Rh6 (green).
(G) tdTomato fluorescence (red).
(H) Merge of F and G.
(I) tdTomato fluorescence superimposed on DIC image of the labellum. I-type sensilla are labelled.
(J) Staining rh6G mutant labellum with anti-Rh6.
(K—M) Staining a rh6+ labellum expressing UAS-tdTomato under the control of the Gr5a-Gal4, which labels sugar GRNs. Anti-Rh6 staining and tdTomato fluorescence are shown.
(K) Anti-Rh6 (green).
(L) tdTomato fluorescence (red).
(M) Merge of K and L.
(N—P) Co-staining a control labellum with anti-Rh6 and anti-DsRed, which labels MSNs (LexAop-rCD2::RFP/+ expressed under the control of the nompC-LexA/+).
(N) Anti-Rh6 (green).
(O) Anti-DsRed (red).
(P) Merge of N and O.