Extended Data Fig. 10: Remote memory recall was inhibited by chemogenetic silencing of mPFC neurons active during remote but not recent memory recall.

(a) Experimental setup. A mixture of AAV-pFos-CreER^T2 and AAV-DIO- hM₄D_i-mCherry (hM₄D_i group) or AAV-DIO-mCherry (mCherry group) was bilaterally injected to the mPFC. mPFC neurons active during memory recall expressed hM₄D_i-mCherry or mCherry. (b) Image showing mPFC neurons expressing hM₄D_i-mCherry (red). Blue fluorescence indicates Nissl stain. (c) Four weeks after CFC, the mice receive a tamoxifen injection (Tam) to label mPFC neurons active during remote memory recall (recall 1) with hM₄D_i-mCherry or mCherry. A week after the recall 1 session, the mice received a CNO injection and were tested for memory recall in the same context (recall 2). (d) Summary plots showing the average freezing time during recall 1 and recall 2 sessions in the hM₄D_i (7 mice) and mCherry groups (8 mice). Repeated measures two-way ANOVA with post hoc comparisons. (e) Two days after CFC, the mice receive a tamoxifen injection to label mPFC neurons active during recent memory recall (recall 1) with hM₄D_i-mCherry or mCherry. Four weeks after recall 1 session, the mice received a CNO injection and were tested for remote memory recall in the same context (recall 2). (f) Summary plots showing the average freezing time during recall 1 and recall 2 sessions in the hM₄D_i (10 mice) and mCherry groups (9 mice). Repeated measures two-way ANOVA with post hoc comparisons. Data are presented as the mean ± SEM.

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