Journal: Molecular Psychiatry

Article Title: Shank3 related oligodendrocyte alterations in autism are restored by Erk pathway inhibition

doi: 10.1038/s41380-025-03333-1

Figure Lengend Snippet: Coronal brain sections from Shank3 WT and KO mice of corpus callosum (CC) have been used for all analyses. A Brain sections from P7 WT and Shank3KO mice stained for MBP and MBP RNA scope. MBP protein and RNA intensity were normalized to mean value. MBP positive cells were shown in the percentage of DAPI. Mean ± SD. Student’s Unpaired t test, p** < 0.01, n = 3 animals. Scale bar = 200 μm. B Immunostaining for MBP and NFH SMI32 and analysis of myelinating and non-myelinating oligodendrocytes in P7 CC. Myelinating and non-myelinating cells were shown in the percentage of total MBP positive cells. Mean ± SD. Student’s Unpaired t test, p**** < 0.0001, n = 3 animals. Scale bar = 100 μm. C P7, P21 and P140 brain section were stained for Olig2 and CC1. Cell number of Olig2 + CC1+ cells were analysed in CC. Data were shown in the percentage of DAPI and Olig2 respectively. Mean ± SD. Student’s Unpaired t test, p* < 0.05, n = 3 animals. Scale bar = 20 μm. D P7 brain section were stained for Ki67, Iba1 and GFAP and proliferation of Ki67+cells was analysed in medial and lateral CC separately. Ki67 positive cells were shown in the percentage of DAPI. Mean ± SD. Student’s Unpaired t test, p* < 0.05, n = 3 animals. Scale bar = 20 μm.

Article Snippet: CC1 (ms) , Merck , Germany , MABC200 , IHC , 1:500 , 10% DS + 5% FBS + 0.3% Triton in PBS-/-.

Techniques: Staining, RNAscope, Immunostaining

Journal: Journal of Cell Science

Article Title: An acentrosomal aster with atypical microtubule polarity recruits cytokinesis signals to its center in Xenopus egg extracts

doi: 10.1242/jcs.263766

Figure Lengend Snippet: Formation of the noncanonical aster requires the microtubule motor MKLP2 and Aurora kinase B activity. (A) Confocal images of microtubule organization in control extracts and extracts with 100 µM MKLP2 inhibitor paprotrain. n =4. Each image is a maximum-intensity projection of nine confocal planes spanning 24 μm of depth. (B) Confocal images of microtubule dynamics in control extracts (top row) and extracts with 40 µM Aurora kinase B inhibitor barasertib (bottom row). n =6. Each image is a maximum-intensity projection of nine confocal planes spanning 16 µm of depth. For both A and B, imaging started at an arbitrary time point when asters had just begun to form in the untreated extracts. (C) Widefield epifluorescence images of microtubule organization in control extracts and extracts with 100 µM kinesin Eg5 inhibitor STLC. n =10. (D) Confocal images of microtubule organization in control extracts and extracts with 2 µM dynein inhibitor GST–p150-CC1. n =6. (E) Widefield epifluorescence images of microtubule and ER organization in control extracts and extracts with 0.68 µM GST–p150-CC1. n =2.

Article Snippet: The chicken DCTN1 p150Glued AA 217–548 (CC1) was from the plasmid pVEX-CC1 (Addgene plasmid 74170; http://n2t.net/addgene:74170 ; RRID:Addgene_74170; deposited by Trina Schroer).

Techniques: Activity Assay, Control, Imaging

Journal: Journal of Cell Science

Article Title: An acentrosomal aster with atypical microtubule polarity recruits cytokinesis signals to its center in Xenopus egg extracts

doi: 10.1242/jcs.263766

Figure Lengend Snippet: Noncanonical asters can merge. (A) Confocal time-lapse montage of microtubule and EB1–GFP dynamics in egg extracts, showing that the centers of two noncanonical asters merged with each another spontaneously, and that the EB1–GFP-enriched regions at the centers also merged. Each image is a maximum-intensity projection of four confocal planes spanning 6 µm of depth. Imaging started at an arbitrary time point after the asters had formed but had not merged. The plot below each image is the fluorescence intensity profile along a 1.65 µm thick, 22.8 µm long line segment (yellow dashed rectangle) that starts at the bottom left and ends at the top right. For each point on the curve in the plot, the horizontal coordinate is the distance from the start of the line segment, and the vertical coordinate is the average fluorescence intensity of the pixels across the width of the line segment at that distance (a.u., arbitrary units). The black arrows indicate intensity peaks for microtubule (second row) and EB1–GFP (fourth row) fluorescence at the aster centers. n =9. (B) Confocal images of microtubules in control and GST–p150-CC1-treated extracts, showing that noncanonical asters still merged when dynein was inhibited by 2 µM GST–p150-CC1. n =2.

Article Snippet: The chicken DCTN1 p150Glued AA 217–548 (CC1) was from the plasmid pVEX-CC1 (Addgene plasmid 74170; http://n2t.net/addgene:74170 ; RRID:Addgene_74170; deposited by Trina Schroer).

Techniques: Imaging, Fluorescence, Control