Extended Data Fig. 6: Induction of new myelin formation preserves remote fear memory recall.
From: Preservation of a remote fear memory requires new myelin formation
a, Extended experimental timeline for cohort represented in Fig. 7a,e. b, Freezing responses in the conditioning context; two-way ANOVA (F4,108 = 5.884, p = 0.0003) with Sidak’s post hoc tests for conditioning (difference: 0.8941 ± 4.116, 95% CI: -9.860 to 11.65, p = 0.9928), 24 hour (difference: -0.8004 ± 4.116, 95% CI: -11.55 to 9.954, p = 0.9948), 7 day (difference: -11.91 ± 4.116, 95% CI: -22.67 to -1.160, p = 0.0221), 21 day (difference: -16.77 ± 4.116, 95% CI: -27.52 to -6.013, p = 0.004), and 30 day (difference: -17.49 ± 4.116, 95% CI: -16.00 to -0.3832, p = 0.0002), n = 15 vehicle and 14 clemastine mice. c, Freezing responses in the similar context; two-way ANOVA (F1,27 = 8.277, p = 0.0077) with Sidak’s post hoc tests for 7 day (difference: 0.4158 ± 3.894, 95% CI: -7.391 to 8.222, p = 0.9154) and 21 day (difference: -8.190 ± 3.894, 95% CI: -28.24 to -6.733, p = 0.401), or across days within vehicle- (difference: -5.845 ± 2.078, 95% CI: -10.77 to -0.9248, p = 0.018) and clemastine-treated (difference: -14.45 ± 2.151, 95% CI: -19.54 to -9.357, p < 0.0001) animals, n = 15 vehicle and 14 clemastine mice. d, Experiment assessing the effects of continuous vehicle injections/handling, compared against home cage animals; two-way ANOVA (F4,36 = 9.926, p < 0.0001) with Sidak’s post hoc tests for conditioning (difference: -0.6013 ± 5.615, 95% CI: -15.66 to 14.46, p > 0.9999), 24 hour (difference: 0.2373 ± 5.615, 95% CI: -14.82 to 15.30, p > 0.9999), 7 day (difference: 22.20 ± 5.615, 95% CI: 7.137 to 37.25, p = 0.0014), 21 day (difference: 31.96 ± 5.615, 95% CI: 16.91 to 47.02, p < 0.0001), and 30 day (difference: 26.78 ± 5.615, 95% CI: 11.72 to 41.83, p < 0.0001), n = 5 mice per treatment group. e, Expanded quantification of Fos+ cell density following 30-day retrieval sessions; unpaired two-tailed t-tests, PL (difference: 279.0 ± 43.19, 95% CI: 188.6 to 369.4, t19 = 6.459, p < 0.0001), IL (difference: 198.6 ± 68.81, 95% CI: 49.90 to 347.2, t19 = 2.886, p = 0.274), ACC (difference: 127.4 ± 51.52, 95% CI: 18.66 to 236.1, t19 = 2.472, p = 0.0069), BA (difference: 86.21 ± 19.27, 95% CI: 45.87 to 126.5, t19 = 4.473, p = 0.0044), LA (difference: 128.6 ± 44.43, 95% CI: 35.89 to 221.2, t19 = 2.894, p = 0.009), DG (difference: 268.7 ± 56.69, 95% CI: 150.0 to 387.3, t19 = 4.739, (p < 0.0001), CA3 (difference: 270.5 ± 77.34, 95% CI: 109.2 to 431.8, t19 = 3.497, p =0.0023), NR (difference: 110.7 ± 34.65, 95% CI: 37.59 to 183.8, t19 = 3.195, p = 0.0026), vDG (difference: 153.9 ± 34.77, 95% CI: 80.51 to 227.2, t19 = 4.426, p = 0.0013), vCA3 (difference: 139.2 ± 36.88, 95% CI: 61.42 to 217.0, t19 = 3.775, p = 0.0019), vPAG (difference: 193.0 ± 48.67, 95% CI: 90.28 to 295.6, t19 = 3.965, p = 0.0017), SSC (difference: 79.25 ± 55.73, 95% CI: -38.33 to 196.8, t19 = 1.422, p = 0.1731), n = 13 vehicle and 8 clemastine mice. f, Within-treatment group comparisons of Fos+ cell density between the PL and IL; unpaired two-tailed t-tests, vehicle (difference: 80.43 ± 48.18, 95% CI: -20.78 to 181.6, t18 = 1.67, p = 0.3657), n = 13 PL- and 7 IL-sampled mice, clemastine (difference: -24.05 ± 65.62, 95% CI: -162.5 to 114.4, t17 = 0.3666, p = 0.7185), n = 8 PL- and 11 IL-sampled mice. Freezing responses for experimental timeline described in (a) for home cage (g), no shock (h), and immediate shock (i) animals; (g) two-way ANOVA (F3,24 = 2.217, p = 0.1122) (h) two-way ANOVA (F4,32 = 2.674, p = 0.0496) (i) two-way ANOVA (F4,32 = 0.7811, p = 0.5496); for g-I, n = 5 animals per condition. j, Extended experimental timeline for clemastine injections in Myrf icKO animals. k, Individual fear expression for vehicle- and clemastine-treated Myrf icKO animals represented in (j); two-way ANOVA (F4,48 =1.357, p = 0.2629), n = 6 vehicle and 8 clemastine mice. l, Quantification of the z-scored mean ΔF/F during pre- and post-bout transitions at 30 days for vehicle- (left) and clemastine-treated (right) animals; paired two-tailed t-tests comparing pre- and post-transition for vehicle (difference: 1.075 ± 0.1560, 95% CI: 0.7685 to 1.382, t78 = 6.894, p < 0.0001) and clemastine (difference: 0.8792 ± 0.1548, 95% CI: 0.5745 to 1.184, t78 = 5.678, p < 0.0001). m, Quantification of the z-scored mean ΔF/F of the pre- and post-bout transition for vehicle- and clemastine-treated animals; unpaired two-tailed t-test (difference: -0.1963 ± 0.2099, 95% CI: -0.6094 to 0.2168, t78 = 0.9352, p = 0.3504). For l-m, n = 10 bouts per animal, 4 animals per treatment group. For box-and-whisker plots, the center, boxes, and whiskers represent the median, interquartile range, and the 10th and 90th percentiles. For dot plots, data are presented as mean ± SEM, with asterisks indicating the following p-value ranges: * ≤ 0.05, ** ≤ 0.01, *** ≤ 0.001, **** ≤ 0.0001.
