Fig. 5. Loss of Nf1 reduces starvation resistance and alters lipid stores and turnover kinetics.

a Quantification of total body triglyceride (TAG) content in Nf1^P1 mutants (n = 9) and w^CS10 controls (n = 10). **p = 0.002 (Mann–Whitney, two-sided). b Quantification of total TAG content of pan-neuronal nSyb-Gal4 > Nf1 RNAi compared to heterozygous controls. p = 0.095 (One-way ANOVA, two-sided; n = 8). c Quantification of total TAG content of PCB-Gal4 > Nf1 RNAi compared to heterozygous controls. p = 0.552 (Kruskal–Wallis, two-sided; n = 24). d Diagram of ¹⁴C sucrose radiolabeling and lipid turnover. e Experimental protocol of ¹⁴C sucrose radiolabeling for lipid turnover analysis. f Quantification of lipid turnover of Nf1^P1 mutants via ¹⁴C sucrose radiolabeling. Radioactivity was measured as counts per minute (CPM) and normalized to fly weight (mg). At time point zero hours (0 h) p = 0.604, at 48-hours (48 h) ***p < 0.001, (Sidak, two-sided; n = 6 for both genotypes at both time points). g Quantification of lipid turnover of PCB-Gal4 > Nf1 RNAi via ¹⁴C sucrose radiolabeling. Radioactivity was measured as counts per minute (CPM) and normalized to fly weight (mg). At time point zero hours (0 h) experimental **p = 0.010 re: UAS-Nf1 RNAi/+, **p = 0.006 re: PCB-Gal4/+. At 48-hours (48 h) experimental p = 0.129 re: UAS-Nf1 RNAi, p = 0.252 re: PCB-Gal4/+. (Sidak, two-sided; n = 12 at 0-h, n = 14 at 48-h). h Lifespan under starvation, comparing Nf1^P1 mutants (n = 78) and w^CS10 controls (n = 80). ***p < 0.001 (X2(1) = 154.0, Mantel–Cox test, two-sided). i Lifespan under starvation, comparing PCB-Gal4 Nf1 knockdown (PCB > Nf1 RNAi) (n = 119) and UAS/+ (n = 120) and Gal4/+ (n = 100) controls. ***p < 0.001 (X2(2) = 281.3, Mantel–Cox test, two-sided). Each “n” in (a–c) represents a sample containing five animals. Each “n” in (f, g) represents a sample containing twenty animals. Box plots—box: 1st to 3rd quartiles, median: line, whiskers: min–max, individual data points: circles. ns: not significant.