(A-D) Heat shock or temperature protocol to induce transgene expression in adult flies (top) and effect on 24-hr memory after spaced training (bottom). HS+ represents heat shock and HS− represents no heat shock. (A) Effect of acute expression of mutant and wild-type csw induced with a heat shock-GAL4 driver and a single heat shock. The memory performance in hsGAL4;UAS-D61Y after heat shock (HS+) was reduced compared with the same line without heat shock (HS−) or control (+/+, UAS-D61Y, UAS-WT, hsGAL4;UAS-WT) groups in both conditions (HS+ or HS−). (B) Effect of acute expression of csw-RNAi (hsGAL4;csw-RNAi) induced with three heat shocks (HS+). Memory was reduced in hsGAL4;csw-RNAi after heat shock (HS+) compared with the same line without heat shock (HS−) or control (+/+, UAS-csw-RNAi, UAS-ctrl-RNAi or hsGAL4;+) groups in both conditions (HS+ or HS−). (C and D) Effect on memory performance of inducible RNAi expression targeted to mushroom bodies using the GAL80ts repressor of GAL4-mediated UAS-csw-RNAi expression, (C) at permissive temperature and (D) at restrictive temperature. Memory was reduced in the GAL80ts;csw-RNAi;OK107 group at restrictive temperature compared with control (+/+, UAS-csw-RNAi, GAL80ts;UAS-ctrl-RANi;OK107), but not at permissive temperature. (E) Effect of the SHP2 inhibitor, NSC-87877 (Inhib [+]) or vehicle (Inhib [−]) on 24-hr memory in flies expressing mutant or wild-type csw in mushroom body neurons. Memory was reduced in the control group (OK107;+) and in flies overexpressing the wild-type csw (WT) by the inhibitor (inhib [+]) compared with both groups fed with vehicle (Inhib [−]), but not in csw gof mutants. Bars, mean ± SEM (A, B and E n=8, and C and D n=5). Asterisks indicate p<0.05.