Extended Figure 1. Behavioral experiment controls.

A) Schematic to test the timecourse of prospective memory-linking (top). Mice underwent Aversive learning and then either 5h, 1d, or 2d later they experienced a Neutral context. The following day, mice were tested in the previously experienced Neutral context. Mice froze significantly more in the Neutral context when the Neutral context occurred within 5h of the Aversive context, compared to when it occurred one day or more after Aversive learning (bottom). Main effect of timepoint (F_2,24 = 3.689, p = 0.04). Post-hoc tests revealed a trend for higher freezing in the 5h timepoint compared to the 1d or 2d timepoints: 1d (t_16.38 = 2.137, p = 0.07), 2d (t_13.45 = 2.38, p = 0.07).

B) Schematic to test the timecourse of retrospective memory-linking (top). Mice experienced a Neutral context, followed by Aversive learning in a separate context 5h, 1d, or 2d later. The day following Aversive learning, they were tested in the previously experienced Neutral context. Mice froze no differently in the Neutral context regardless of how long before Aversive learning the Neutral context was experienced (bottom). No main effect of timepoint (F_2,27 = 0.73, p = 0.49).

C) Schematic of low vs high shock retrospective memory-linking experiment (without calcium imaging as a replication). Mice received a Neutral experienced followed by a low or high shock Aversive learning two days later. In the subsequent 3 days, mice were tested in the Aversive context, and then Neutral and Novel contexts, counterbalanced.

D) Mice froze more in the Aversive context in High Shock vs Low Shock mice (t₁₄ = 5.04, p = 0.00018).

E) High Shock mice exhibited higher freezing in Neutral vs Novel recall, while Low Shock mice did not. A priori post-hoc test: High Shock (t₇ = 2.65, p = 0.033), Low Shock (t₇ = 1.21, p = 0.133).