Functional analysis of Dh44, Dh44R1, and Dh44R2 mutants and their roles in lipid accumulation. (A) Total TAG level (µg TAG/mg fly) measurement in whole body extracts from control (w1118), Dh44Mi, Dh44R1Mi, and Dh44R2Mi mutants (n=5). (B–E) Nile red stains of the fat body from w1118, Dh44Mi, Dh44R1Mi, and Dh44R2Mi respectively. Scale bars represent 50 µm. (F) Area of LDs in each indicated genotype (n=3). (G) The protein (µg protein/mg fly) measurement in the whole-body extracts from control (w1118) and Dh44Mi, Dh44R1Mi, and Dh44R2Mi mutants (n=4). (H) Quantification (qRT-PCR) of lipolytic gene (bmm) expression level in the fat body of w1118, Dh44Mi, Dh44R1Mi, and D44R2Mi flies under sated and starved conditions (n=3). (I) Total TAG level measurement in whole body extracts from the indicated genotypes (n=4). (J–M) Nile red stains of the fat body from (J) w1118, (K) trpγ1 (L) trpγ1,UAS-trpγ1/trpγ1,Dh44R1-GAL4, and (M) trpγ1,UAS-trpγ1/trpγ1,Dh44R2-GAL4. (N) Area of LDs in each indicated genotype (n=3). Scale bar represent 50 µm. (O-P) Expression of Dh44R2 in the brain and intestine. (O) Expression of Dh44R2 in the subesophageal zone (SEZ) of the brain. Scale bars represent 50 µm. (P) Expression of Dh44R2 in the intestine. Scale bars represent 200 µm in full intestine and 500 µm in magnified form.
Means ±SEMs. Single factor ANOVA with Scheffe’s analysis was used as a post hoc test to compare multiple sets of data. The asterisks indicate significance from control (*P < 0.05, **P < 0.01). Each dot indicates distribution of individual sample value.