Extended Data Fig. 2: Eif2a cKI^Camk2a mice show enhanced protein synthesis and consolidation of contextual and auditory fear memory.

a, b, Immunofluorescent labelling of excitatory neurons shows a decrease in p-eIF2α levels in Eif2a cKI^Camk2a compared to control mice. Two independent experiments showed similar results. c, Short-term contextual memory is not altered in Eif2a cKI^Camk2a mice (F _{2, 40} = 0.97; n = 8, 7, 8). d, Short-term auditory fear memory is not altered in Eif2a cKI^Camk2a mice (F _{2, 42} = 0.46; n = 8, 8, 8). e, Illustration of cannulation site for unilateral injections of puromycin into the CA1 region. f, Experimental design of iSUnSET assay. g, Immunofluorescent images of puromycin in Eif2a^A/AfTg⁺ mice injected with AAV9.Camk2a0.4.Cre.SV40 (Camk2a-Cre) shows enhanced protein synthesis (t _50.43 = 6.08; n = 26, 27, points represent group means). Five independent experiments showed similar results. h, Schematic representation of strong contextual and auditory fear conditioning. i, Contextual memory is enhanced in Eif2a cKI^Camk2a mice following a strong training protocol (F _{2, 44} = 12.97; n = 8, 9, 8). j, Auditory fear memory is enhanced in Eif2a cKI^Camk2a mice following a strong training protocol (F _{2, 42} = 7.64; n = 8, 8, 8). k, In an open field test, all groups spent a similar amount of time in the outer and inner zones (F _{2, 54} = 5.6 × 10⁻¹²; n = 10, 10, 10). Representative heat-map of travelled path in an open field arena. l, The mean total distance moved (in cm) in the open field was similar in all groups (F _{2, 27} = 0.19; n = 10, 10, 10). m, Open field test arena with illustration of outer and inner zones. Data are presented as mean ± s.e.m. in c, d, g, i–l. p-values by two-tailed unpaired t-test with Welch’s correction in g and by two-way ANOVA in i and j followed by Tukey’s multiple comparisons post hoc test are indicated. Points represent individual mice. Scale bars: 20 μm.

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