Fig. 3: Functional and cytological validation of Gnmt, Sardh, and CG5955 KD using DJ694-Gal4. | Nature Communications

Fig. 3: Functional and cytological validation of Gnmt, Sardh, and CG5955 KD using DJ694-Gal4.

From: Time-restricted feeding promotes muscle function through purine cycle and AMPK signaling in Drosophila obesity models

Fig. 3

a Flight performance of female flies with DJ694-Gal4-driven KD of Gnmt, Sardh, or CG5955 at 4 days and 3 weeks of age. Two independent RNAi lines were used for control and each gene. Flight indices plotted as cohorts of 10–20 flies from independent RNAi lines were indicated with the symbol circle or triangle. # of cohorts per RNAi line per condition = 3; # of flies per genotype per age group = 86–130. Details of precise N are in Source Data. Mean ± SEM. One-way ANOVA with Sidak post hoc tests. b Fluorescence images of the IFMs from 3-week-old female flies with DJ694-Gal4-driven KD of Gnmt, Sardh, and CG5955 upon staining with phalloidin (green) and Nile Red (red puncta). More accumulation of lipid (arrows) and actin-containing myofibrillar disorganization (asterisks) was detected in IFMs upon KD of Gnmt, Sardh, and CG5955, compared with age-matched control. The scale bar is 20 µm. c, d Intramuscular lipid quantification (lipid droplet size (c) and density (d)). N = 9 from three flies’ IFM per genotype. Mean ± SEM. One-way ANOVA with Sidak post hoc tests. e Flight performance of 5-week-old female flies with DJ694-Gal4-driven KD of Gnmt, Sardh, and CG5955 under ALF and TRF. Two independent lines were tested. Flight indices were plotted as cohorts of 10–20 flies. Results from independent RNAi lines were indicated with symbol circle or triangle. # of cohorts per RNAi line per condition = 3–4; # of flies per genotype = 67–98. Details of precise N are in Source Data. Mean ± SEM. Two-way ANOVA with Sidak post hoc tests. f Relative expression of Gnmt in 3-week-old female flies with Gnmt overexpression using DJ694 driver. Experimental flies were fed with RD or HFD. N = 3 biological replicates. Mean ± SEM. Two-sided unpaired t-tests. g Flight performance in 3-week-old female flies with Gnmt overexpression driven by DJ694. Experimental flies were fed with RD or HFD. Flight indices were plotted as cohorts of 10–20 flies. # of cohorts per condition = 3–5; # of flies per genotype per age = 40–100. Details of precise N are in Source Data. Mean ± SEM. Two-sided unpaired t-tests. Source data are provided as a Source Data file.

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