The flies used to obtain data for this figure were uas-armRNAi/+; P{MB-GeneSwitch}12-1,uas-dcr2/MB-GCamP and were trained using 5X spaced forward conditioning.

(A) Performance of flies with and without RU486 feeding at 24 hr after conditioning using benzaldehyde (Ben) as the CS+ and 3-octanol (Oct) as the CS−. The RU-fed group showed a significantly reduced performance index compared with the unfed group of the same genotype (n = 8, ∗p < 0.05).

(B–D) A small fraction of the behaviorally trained flies were removed prior to testing and used for functional imaging.

(B) Average peak calcium responses in the α branch of the α/β MB neurons to the CS+ (Ben) and CS− (Oct) at 24 hr after conditioning with benzaldehyde as the CS+. There was a significant difference in response to the CS+ between the RU-fed and no-RU groups (∗p < 0.05). There was no significant difference in response to the CS− between the RU-fed and no-RU groups (n = 9 and 10 for no-RU and RU-fed groups, respectively).

(C) Group time course for calcium responses in the α branch of the α/β MB neurons during presentation of the CS+ (Ben) at 24 hr after conditioning with Ben as the CS+ (n = 9 and 10 for no-RU and RU-fed groups, respectively). The graph was made using the data from the same flies used for the bar graph in (B).

(D) Group time course for calcium responses in the α branch of the α/β MB neurons across time during presentation of the CS− (Oct) at 24 hr after conditioning with Ben as the CS+ (n = 9 and 10 for no-RU and RU-fed groups, respectively). The graph was made using the data from the same flies used for the bar graph in (B).

(E) Performance of flies with and without RU486 feeding at 24 hr after conditioning using Oct as the CS+ and Ben as the CS−. The RU-fed group showed a significantly reduced performance index compared with the no-RU group of the same genotype (n = 8, ∗p < 0.05).

(F–H) A small fraction of the behaviorally trained flies were removed prior to testing and used for functional imaging.

(F) Average peak calcium responses in the α branch of the α/β MB neurons to the CS+ (Oct) and CS− (Ben) at 24 hr after conditioning with octanol as the CS+. There was a significant difference in response to the CS+ between the RU-fed and no-RU groups (∗p < 0.05). There was no significant difference in response to the CS− between the RU-fed and no-RU groups (n = 9 and 8 for no-RU and RU-fed groups, respectively).

(G) Group time course for calcium responses in the α branch of the α/β MB neurons across time during the presentation of the CS+ (Oct) at 24 hr after conditioning with octanol as the CS+ (n = 9 and 8 for no-RU and RU-fed groups, respectively). The graph was made using the data from the same flies used for the bar graph in (F).

(H) Group time course for calcium responses in the α branch of the α/β MB neurons across time during the presentation of the CS− (Ben) at 24 hr after conditioning with octanol as the CS+ (n = 9 and 8 for no-RU and RU-fed groups, respectively). The graph was made using the data from the same flies used for the bar graph in (F). Error bars are the SEM.