Extended Data Fig. 8: Validation of Robust Activity Marking (RAM) technique for silencing engram neurons.
From: The astrocytic ensemble acts as a multiday trace to stabilize memory
a, Schematic showing the injection of RAM-hM4Di-mCherry or RAM-NLS-mKate2 into the amygdala. Mice underwent fear conditioning two weeks later followed by i.p. CNO (1 mg/kg) injections the day after. Brain samples were intracardially fixed for immunohistochemistry analysis. b, Graphs showing the percentage of RAM-expressed neurons in LA/B (left), co-expression of c-Fos and RAM (center), and overall c-Fos+ neurons (right). mKate2: n = 15 slices, hM4Di: n = 14 slices, 5 mice per group. Mann-Whitney test. ****p < 0.0001. c, Representative images (from mKate2: n = 15 slices, hM4Di: n = 14 slices, 5 mice/group) showing expression of neuronal marker NeuN and RAM-tagged engram neurons (indicated by mCherry signals) in b. Overlapping c-Fos-expressing engram neurons were fewer in the RAM-hM4Di group. Data are mean ± SEM. Sample sizes and replicates in Supplementary Table 1.
