Extended Data Fig. 1: , related to Fig. 1: Control experiments for FA mitochondrial import and β-oxidation in MB neurons to sustain memory formed after massed training.
From: Neuronal fatty acid oxidation fuels memory after intensive learning in Drosophila
Extended Data Fig. 1a. When CPT1 RNAi expression was not induced, memory after massed training was normal (n = 12, F2,33 = 0.30, P = 0.743). b. A second non-overlapping RNAi targeting CPT1 (CPT1 RNAi KK100935) was used to confirm the specific defect in memory after massed training. Inhibition of CPT1 expression in adult MB neurons using CPT1 RNAi KK100935 impaired memory after massed training (n = 20, F2,57 = 8.91, P = 0.0004), whereas memory after spaced training was normal (n = 18, F2,51 = 0.43, P = 0.656). Non-induced flies showed no memory defect after massed training (n = 18, F2,51 = 0.61, P = 0.546). c. When MTPα RNAi expression was not induced, memory after massed training was normal (n = 10, F2,27 = 0.21, P = 0.816). d. A second non-overlapping RNAi targeting MTPα (MTPα RNAi GD11299) was used to confirm the specific defect in memory after massed training. In order to increase RNAi efficiency, Dicer2 expression was induced together with the RNAi expression in adult MB neurons using the Tub-Gal80ts; UAS-Dcr2, VT30559 line. Inhibition of MTPα expression in adult MB neurons using this RNAi impaired memory after massed training (n = 18, F2,51 = 7.30, P = 0.0016), whereas memory after spaced training was normal (n = 14, F2,39 = 1.21, P = 0.309). Non-induced flies showed no memory defect after massed training (n = 12, F2,33 = 2.62, P = 0.088). e. When HAD1 RNAi expression was not induced, memory after massed training was normal (n = 13-15, F2,39 = 1.63, P = 0.209). f. Because only one RNAi line targeting Had1 was available, we used a HAD1 null mutant to confirm HAD1 function in memory formed after massed training, albeit without cell-type specificity. Since HAD1nl is in a yw background, we used heterozygous +/yw flies as a control for the +/HAD1nl experimental group. Heterozygous +/HAD1nl flies had a memory defect after massed training as compared to control +/yw flies (n = 12, t22 = 2.97, P = 0.0071), whereas memory after spaced training was normal (n = 16, t30 = 1.23, P = 0.229). Data on barplots are expressed as mean ± s.e.m. with dots as individual values, and analyzed by ANOVA with post hoc testing by Newman–Keuls pairwise comparisons test (a-e) or by two-sided unpaired t-test (f). Genotype sample sizes are listed in the legend in order of bar appearance. Significance level of a two-sided unpaired t-test (f) or the Newman–Keuls pairwise comparison between the genotype of interest and the genotypic controls (a-e) following one-way ANOVA: **P < 0.01, ***P < 0.001, NS: not significant.
