module plug  (Abaqus Inc)

Journal: Nature Communications

Article Title: Elevated levels of FMRP-target MAP1B impair human and mouse neuronal development and mouse social behaviors via autophagy pathway

doi: 10.1038/s41467-023-39337-0

Figure Lengend Snippet: a A schematics showing 5q13.2 triplication in the ASD patient. Blue boxes: genes affected. b mRNA levels in fibroblasts of the ASD patient and his mother (Ctrl). Two-tailed, unpaired Student’s t -test. MAP1B : p < 0.0001; MRP27 : p < 0.0001; PTCD2 : p = 0.0002; ZNF366 : p = 0.0016. n = 3 technical replicates, N = 1. c Experimental scheme for analyzing ASD patient neurons. d Western blot analysis of MAP1B levels. GAPDH: loading control. One-way ANOVA with Dunnett post hoc tests, p = 0.0284. e Representative confocal images (from three independent experiments) and Neurolucida traces of GFP+ neurons. Scale bar, 10 μm. f Sholl analysis of ASD patient neurons with ( shMAP1B ) or without ( shNC ) MAP1B knockdown. MANOVA, F (1,140) = 12.001, p < 0.001. g Total dendritic length. Two-tailed, unpaired Student’s t -test, p < 0.0001. For ( f ) and ( g ), shNC : n = 70 neurons, shMAP1B : n = 72 neurons from three differentiations, N = 1. h Representative raster plots. i Neuronal mean firing rate. Two-tailed, unpaired Student’s t -test, p < 0.0001. Ctrl: n = 38 wells, MAP1B-EE: n = 25 wells from 3 differentiations, N = 1. j Number of bursts. Two-tailed, unpaired Student’s t -test, p = 0.0010. Only the wells with bursting activity were analyzed for burst-related parameters, shNC : n = 23 wells, shMAP1B : n = 20 wells from three differentiations, N = 1. k Representative traces showing current injection-evoked action potentials. l Rheobase current threshold. Two-tailed, unpaired Student’s t -test, p = 0.0160. shNC : n = 6 neurons, shMAP1B : n = 6 neurons from at least four differentiations, N = 1. m Representative traces of mEPSCs. n , o Cumulative probability of interevent intervals and amplitudes of mEPSCs. Two-sided, Mann–Whitney Rank Sum test, p < 0.0001. (Inset: mean mEPSC frequency, p = 0.0472 and amplitude, p < 0.0001. Two-sided, unpaired Student’s t -test). shNC : n = 30 neurons, shMAP1B : n = 28 neurons from three differentiations, N = 1. Error bars reflect mean ± s.e.m. Source data are provided as a Source Data file.

Article Snippet: The dendritic morphology of a single neuron was analyzed by Neurolucida software with a 3D module plug-in (MicroBrightField, Inc. Williston, VI, http://www.mbfbioscience.com/ ).

Techniques: Two Tailed Test, Western Blot, Control, Knockdown, Activity Assay, Injection, MANN-WHITNEY

Journal: Nature Communications

Article Title: Elevated levels of FMRP-target MAP1B impair human and mouse neuronal development and mouse social behaviors via autophagy pathway

doi: 10.1038/s41467-023-39337-0

Figure Lengend Snippet: a Sholl analysis of FXS2 neurons with MAP1B knockdown. F (1,63) = 20.697, p < 0.001, shNC n = 30 neurons, shMAP1B n = 35 neurons from three independent neuronal differentiations, N = 1. b Quantification of total dendritic length. Two-tailed, unpaired Student’s t -test, p < 0.0001, shNC n = 30 neurons, shMAP1B n = 35 neurons from three independent neuronal differentiations, N = 1. c Neuronal mean firing rate. Two-tailed, unpaired Student’s t -test, p = 0.0032. C t rl2: n = 10 individual wells, FXS2: n = 24 individual wells from three independent neuronal differentiations, N = 1. d Mean firing rate in FXS2 neurons with MAP1B knockdown or rapamycin treatment. One-way ANOVA with Dunnett post hoc tests, shNC +Veh vs. shMAP1B +Veh: p = 0.0165; shNC +Veh vs. shNC +Rap: p = 0.0027. shNC Veh: n = 24 individual wells, shMAP1B Veh: n = 24 individual wells, shNC Rap: n = 22 wells from three independent neuronal differentiations, N = 1. e Representative confocal images of neurons expressing shRNA-mCherry (red), LC3 (white) in lentivirus-infected ex vivo human cortical slices. Scale bars: 20 μm. f LC3 intensity in mCherry+ neurons in human cortical slices. Two-way ANOVA with two-sided Bonferroni post hoc analysis for multiple comparisons: shNC+ Veh vs. shFMR1 +Veh: p = 0.0476; shFMR1 +Veh vs. shFMR1 +Rap: p = 0.0332. N = 3 individual cortices. g Representative confocal images (from three independent experiments) and Neurolucida traces of mCherry+ neurons. Scale bar, 10 μm. h Sholl analysis. MANOVA, shNC +Veh vs. shFMR1 Veh: F (1,91) = 51.474, p < 0.001; shFMR1 +Veh vs. shFMR1 Rap: F (1,108) = 54.811, p < 0.001. i Total dendritic length of LV- shFMR1 or LV- shNC -infected neurons treated with Veh or Rap. Two-way ANOVA with two-sided Bonferroni post hoc analysis for multiple comparisons, shNC+ Veh vs. shFMR1 +Veh: p < 0.0001; shFMR1 +Veh vs. shFMR1 +Rap: p < 0.0001. For all data shown in ( g , h ) , n = 49 ( shNC+ Veh) neurons; n = 54 neurons ( shNC +Rap); n = 44 neurons ( shFMR1 +Veh); n = 66 neurons ( shFMR1 +Rap) from N = 3 individual cortices. All error bars reflect mean ± s.e.m. Source data are provided as a Source Data file.

Article Snippet: The dendritic morphology of a single neuron was analyzed by Neurolucida software with a 3D module plug-in (MicroBrightField, Inc. Williston, VI, http://www.mbfbioscience.com/ ).

Techniques: Knockdown, Two Tailed Test, Expressing, shRNA, Infection, Ex Vivo