<i>dbr</i> RNAi expression in the adult MBs impairs LTM formation.

<p>(A) <i>dbr</i> mRNA expression analyses. Total RNA was extracted from fly heads, submitted to DNase treatment, and further reverse transcribed with oligo(dT) primers. Resulting cDNA was quantified by PCR using tubulin (<i>Tub</i>) expression as a reference. Results are shown as ratios relative to the values observed for <i>elav/+</i> (left panel, <i>t</i> test, <i>p</i><0.01, n≥5) and wild-type (+, right panel, <i>t</i> test, <i>p</i>>0.05, n≥4) control flies. (B–I) Behavioral analyses. To silence <i>dbr</i> expression in the adult MBs, flies were fed with RU for 48 hrs prior to conditioning and in (G–I), also for 24 hrs before testing. (B) Shock reactivity (ANOVA, left panel, <i>p</i> = 0.604, <i>n</i> = 9; right panel, <i>p</i> = 0.1049, <i>n</i>≥8). (C) Olfactory acuity for octanol (ANOVA, left panel, <i>p</i> = 0.8099, <i>n</i> = 10; right panel, <i>p</i> = 0.0891, <i>n</i>≥8). (D) Olfactory acuity for methylcyclohexanol (ANOVA, left panel <i>p</i> = 0.9488, <i>n</i>≥8; right panel, <i>p</i> = 0.1633, <i>n</i>≥8). (E) Learning analysis after a single conditioning cycle (ANOVA, <i>p</i> = 0.5396, <i>n</i> = 8). (F) STM analysis 2 hrs after a single conditioning cycle (ANOVA, left panel, <i>p</i> = 0.6264, <i>n</i>≥10; right panel, <i>p</i> = 0.0472, <i>n</i>≥8, the Newman-Keuls post-test is only significant for the <i>MB-Sw/RNAi-B1</i> vs <i>MB-Sw/+</i> pair, <i>q</i> = 4.474). (G) ARM analysis 24 hrs after five massed conditioning cycles (ANOVA, left panel, <i>p</i> = 0.5088, <i>n</i> = 16; right panel, <i>p</i> = 0.7635, <i>n</i>≥13). (H) LTM analysis 24 hrs after five spaced conditioning cycles (ANOVA, left panel, <i>p</i><0.0001, <i>n</i>≥21; right panel, <i>p</i><0.0001, <i>n</i>≥14). (I) In the absence of RU, LTM is not affected. Flies were either raised on regular medium (hatched bars) or fed with RU for 48 hrs prior to training, and until testing (+RU) (gray bars). Scores were measured 24 hrs after a five-spaced cycle conditioning (<i>t</i> test: left panel, <i>MB-Sw/RNAi-A</i> vs <i>MB-Sw/+</i>, <i>p</i> = 0.8335, <i>MB-Sw/RNAi-A</i> vs <i>MB-Sw/RNAi-A</i> (+RU), <i>p</i> = 0.0222, <i>n</i> = 12; right panel, <i>MB-Sw/RNAi-B1</i> vs <i>MB-Sw/+</i>, <i>p</i> = 0.4432, <i>MB-Sw/RNAi-B1</i> vs <i>MB-Sw/RNAi-B1</i> (+RU), <i>p</i> = 0.0005, <i>n</i>≥8 ; <i>MB-Sw/RNAi-B5</i> vs <i>MB-Sw/+</i>, <i>p</i> = 0.5989, <i>MB-Sw/RNAi-B5</i> vs <i>MB-Sw/RNAi-B5</i> (+RU), <i>p</i> = 0.0015, <i>n</i>≥8). Bars indicate Mean ± SEM, ns, not significant.