EMBO/sd-character-level-ner huggingface dataset:
Figure 3A. Top row: MG132 arrested HeLa cells stably expressing tubulin-GFP (green) and immunostained for endogenous PRC1 (AlexaFluor555 shown in magenta) in 5 individual z-images. Enlargements of the boxed region (top: merge, middle: GFP, bottom: AlexaFluor555) below the first image show the bridging fiber determined as the line connecting two ends of the sister k-fibers in green channel and localization of endogenous PRC1 (magenta). Corresponding maximum projections are shown to the right (max z, left: merge, middle: GFP, right: AlexaFluor555).B. Quantification of tubulin-GFP bridging fibers (green bar) immunostained for endogenous PRC1 (magenta bar). C. Images show pole-to-pole tracking (white curve) of tubulin signal (top, green), which was used to measure the endogenous PRC1 signal along the same contour (middle, magenta), and the corresponding scheme (bottom) from a HeLa cell expressing tubulin-GFP and immunostained for PRC1. Images of the same spindle without (left) and with the tracked contour (right) are shown. The graph shows normalized pole-to-pole intensity profiles (each intensity profile was scaled so that one pole is at x=0 and the other at x=1) of endogenous PRC1 (grey lines) acquired in tubulin-GFP HeLa cells immunostained for PRC1. Black line shows the mean value. D. Images show pole-to-pole tracking (white curve) of PRC1-GFP signal (top, merge; middle, GFP; bottom, scheme) from a HeLa cell expressing PRC1-GFP and mRFP-CENP-B. Images of the same spindle without (left) and with the tracked contour (right) are shown. The graph shows normalized pole-to-pole intensity profiles (legend as in C) of PRC1-GFP acquired in HeLa cells expressing PRC1-GFP and mRFP-CENP-B. E. Example of length measurement of the immunostained PRC1 signal (LPRC1) in HeLa cell expressing tubulin-GFP and immunostained for PRC1. F. Comparison of the length LPRC1 between PRC1-GFP signal (green, corresponds to the PRC1 fused to GFP from a PRC1-GFP cell line) and PRC1-AlexaFluor555 (magenta, corresponds to the endogenous immunostained PRC1) (p=0.10). G. Cross section signal intensity of the immunostained endogenous PRC1 (magenta line) from a HeLa cell expressing tubulin-GFP and immunostained for PRC1. Area under the peak is defined as Icross, measured at the position of the blue line as in scheme. Horizontal lines mark the background signal (black line), vertical lines delimit the area (grey) where the signal was measured.
Interaction of PTP-MEG2 and tyrosine-phosphorylated NSF in the medulla and the crystal structure of the PTP-MEG2/phospho-NSF-segment complex.Figure 2(A). NSF was phosphorylated after stimulation with AngII or KCl. Adrenal medulla cells were stimulated with 100 nM AngII or 70 mM KCl for 5 min and lysed. NSF was immunoprecipitated with a specific NSF antibody coated with Protein A/G beads. A pan-phospho-tyrosine antibody pY20 was used in Western blot to detect the tyrosine-phosphorylated NSF in the adrenal medulla under different conditions. (B). NSF was co-localized with PTP-MEG2 in the adrenal medulla. After stimulation with 100 nM AngII, NSF and PTP-MEG2 in the adrenal medulla were visualized with immunofluorescence. White arrow stands for co-localization of the PTP-MEG2 and NSF. (C). Analysis of PTP-MEG2 and NSF fluorescence intensities by Pearson's correlation analysis. The Pearson's correlation coefficient was 0.7. (D). Phosphorylated NSF interacted with PTP-MEG2. PC12 cells were transfected with FLAG-NSF. After stimulation of the cells with 100nM AngII in the presence or absence of 100μM H2O2 for 5 minutes (H2O2 was exploited to increase the overall tyrosine phosphorylation of the cellular proteins), the potential PTP-MEG2 substrate in cell lysates was pulled down with GST-PTP-MEG2-D470A or GST control. (E). The overall structure of PTP-MEG2-C515A/D470A in complex with the NSF-pY83 phospho-segment. (F). The 2Fo-Fc annealing omit map (contoured at 1.0σ) around the NSF-pY83 phospho-segment. The P-loop was highlighted in blue. (G). Plot of distance RMSDs of individual residues between the crystal structures of the PTP-MEG2/NSF-pY83 phospho-peptide complex (PDB: 6KZQ) and the PTP-MEG2 native protein (PDB: 2PA5). (H). Superposition of the PTP-MEG2/NSF-pY83 phospho-peptide complex structure (red) on the PTP-MEG2 native protein structure (PDB: 2PA5, blue). The structural rearrangement of the WPD loop and β3-loop-β4 is highlighted. (I, J). The closure of the WPD loop and corresponding conformational changes in the inactive state (I) and the active state (J) of PTP-MEG2. The rotation of R521 leads to the movement of W468 and a corresponding 7 Å movement of the WPD loop. (K, L). The structural rearrangement of β3-loop-β4 of PTP-MEG2 in the active state (L) compared with the inactive state (K). The disruption of the salt bridge between E406 and R521 contributed to the new conformational state of the N-terminal of β3-loop-β4. (M, N). The conformational change of the P-loop of the inactive state (M) and the active state of PTP-MEG2 in response to NSF-pY83 segment binding (N). The disruption of the charge interaction between R409 and D335 resulted in the movement of the main chain from G408 to K411.
Figure 1A. Lysates from 293T cells transfected with the indicated DNA constructs were incubated with 1 mM Mg++ plus 0.1 mM GTPγS, GDP or 2 mM EDTA and immunoprecipitated with anti-Myc antibody. Rab18-HA co-immunoprecipitated with Myc-TRAPPC9 when it was nucleotide free (+EDTA). B. A mutant of Rab18 unable to bind nucleotide, N122I, was co-immunoprecipitated with Myc-TRAPPC9 by anti-Myc antibody. C. HA-tagged Rab1 and Rab18, but not Rab2, could co-immunoprecipitate endogenous TRAPPII complex but not TRAPPIII complex. D. GST-Rab18 could pull down endogenous TRAPPII complex in an in vitro binding assay. GST-Rab18 was pre-loaded with GTPγS or GDP, or stripped off nucleotide by EDTA. Lysates from 293T cells were subjected to GST-Rab18 pull-down. The presence of various TRAPP subunits was detected by immunoblotting. TRAPPII, but not TRAPPIII complex was co-isolated with GST-Rab18, most strongly with Rab18 in nucleotide-free state.
Figure 5(A) Genome browser tracks showing H3K27me3 (top) and H4K20me1 (bottom) accumulation at a gene silenced rapidly (Rnf12) or more slowly (Pgk1). Allele-specific tracks were overlaid (B6 in red and Cast in blue). Note strong H4K20me1 bi-allelic pre-marking at gene bodies. (B) Average H3K27me3 (top) and H4K20me1 (bottom) enrichment at the B6 allele over initially active or inactive genes ± 30kb at the X chromosome. Shown is data for all time points. Data information: TSS : transcription start site ; TES : transcript end site. (C) Average H4K20me1 enrichment at the B6 allele over early and late silenced genes ± 30kb at the X chromosome. Shown is data for all time points. Data information: TSS : transcription start site ; TES : transcript end site.
Figure 4a, Representative flow cytometry data of CD69 and Celltrace violet (CTV) staining on gated living CD8 T cells. The purified CD8 Tn were isolated from young mice and stimulated by different doses of anti-CD3 ab for 72 hours. Enlarged number representing the percentage of the corresponding gate. b, the percentages of proliferating cells among living CD8 singlets (CD8 Tn were from young mice). c, the percentages of CD69+ cells among living CD8 T singlets (CD8 Tn were from young mice). d, Representative flow cytometry data of CD69 and CTV staining on gated living CD8 T cells. The purified CD8 Tn were isolated from 45-wk-old mice and stimulated by different doses of anti-CD3 ab for 72 hours. Enlarged number representing the percentage of the corresponding gate. e, the percentages of proliferating cells among living CD8 singlets (CD8 Tn were from 45-wk-old mice). f, the percentages of CD69+ cells among living CD8 T singlets (CD8 Tn were from 45-wk-old mice).
Figure 1(A) Domain architecture of human HSP27: N-terminal domain (N-term), α-crystallin domain (ACD), and C-terminal region (C-term). The conserved IxI/V motif in the C-terminal region is indicated. Missense, nonsense, and frameshift mutations associated with CMT disease and dHMN are shown. The P182L mutation in the IxI/V motif is blue. Residues with multiple mutations are italicized (e.g. P182L, P182S). The asterisk denotes a nonsense mutation and the X indicates a frameshift mutation. (B) Amino acid sequence alignment of IxI/V motifs and adjacent residues from vertebrate HSP27 orthologues: human (Homo sapiens), mouse (Mus musculus), alligator (Alligator mississippiensis), python (Python bivittatus), frog (Xenopus laevis), shark (Callorhinchus milii), gar (Lepisosteus oculatus), rice fish (Oryzias latipes), and lungfish (Protopterus annectens). (C) Domain boundaries of the ACD used in this study, cHSP27, and residues in the C-terminal IxI/V peptide. (D) Three-dimensional structure of the ACD dimer (black) bound to the IxI/V peptide from the C-terminal region (red). The β-strands of the ACD are numbered. (E) Zoomed-in region from D showing the contacts made between the IxI/V peptide and the ACD for the wild-type peptide. (F) Malate dehydrogenase (MDH; 0.2 µM) was incubated at 40 °C in the absence (black) or presence of 0.5 µM WT HSP27 (red) or 0.5 µM P182L HSP27 (blue). (G) Insulin (40 µM) was incubated at 40 °C in the absence (black) or presence of 20 µM WT HSP27 (red) or 20 µM P182L HSP27 (blue). The y-axes depict the absorbance at 340 nm due to the formation of large aggregates. The solid lines represent the average of three replicates with the filled area reflecting ± one standard deviation.
Figure 3(A) Heat maps representing the genes significantly downregulated or upregulated in HF feeding and HF fecal transplantation at ZT0 and ZT12 (n=4, Cyber-T test p<0.05). (B) Heat map of PPARγ target genes upregulated in both HFD feeding and HF-R at ZT12 (n=4, Cyber-T test p<0.05). (C) KEGG pathway analysis of genes downregulated at ZT12 in both HF feeding and HF-R.(D) KEGG pathway analysis of genes upregulated at ZT12 in both HFD feeding and HF-R. (E) Venn diagram and heatmap of metabolites whose expression increased in both HFD feeding and HF-R at ZT12 (n = 5, Cyber-T test p<0.05).
Figure 5(A-F) TEM tomograms and 3D reconstructions of terminal cells in shibirets embryos. (A) Control. (B-F) Kept at 34ºC for 1 (B, E) or for 2 hours (C, D, F). Boxed regions in (A) and (B) are magnified in (A'') and (B''). (A', B') Reconstructed 3D models. The tube membrane (cyan) is characterized by the electron-dense apical extracellular matrix (aECM). Invaginations with connections that can be traced to the tube membrane are red, invaginations from the basal plasma membrane are green. The basal membrane is light pink. (C-D') shibirets terminal cell after 2 hours at 34ºC. (C') 3D reconstruction, color coding for basal and tube membrane is the same as above. Membrane that is continuous with both the apical and the basal membrane is shown in green, purple and dark blue. (D-D') higher magnification of a region in the same cell at a level where membrane sheets (purple, dark blue) bridge the apical and basal plasma membrane domains. (E-E'') Two focal planes and model of a tomogram from the cell shown in (B) where a basal (pink arrowhead) and an apical invagination (blue) are seen in close proximity (closest distance is marked by cyan and blue arrowheads). (F-F') apical membrane overgrowth regions of the cell shown in (C). The genotype of the embryos was shibirets; btl>KDEL::RFP, Par3::YFP. The cell shown in (C-D) was found and acquired without the CLEM approach.
Figure 3(A and B) Immunoblot analyses of indicated membrane fractions prepared by differential centrifugation. (A and B) Immunoblot analyses of indicated membrane fractions prepared by differential centrifugation. HeLa cells reconstituted with flag-pro-IL-1β and myc-pro-Caspase-1 were treated with 1mM LLOMe for 1h, and lysates were subjected to differential centrifugations at 3,000xg (3k), 25,000xg (25k) and 100,000xg (100k) as depicted in the cartoon. Arrows, two ways the membranes were analyzed - pellets from velocity sedimentation were analyzed in panels B-D, or by further sucrose and OptiPrep density separation of 25k pellets in panels F and G. (C) IL-1β levels in 25k membrane fractions (pellets) from HeLa cells and their CRISPR knockout derivatives (TRIM16KO) that were reconstituted for IL-1β secretion and treated with 1mM LLOMe for 1 h. (D) Quantification of analyses in (B). RI, relative intensity. (E) Whole cell lysates (immunoblots) analyzed in F and G. (F,G) Sucrose density gradient analysis of 25k membranes. Note that mature IL-1β (mIL-1β) is recruited to and co-fractionates with TRIM16+, Sec22b+, and LC3-II+ membranes in TRIM16wt HeLa cells reconstituted with pro-IL-1β and pro-caspase 1, but not in TRIM16KO mutant cells. (H) Confocal images of LC3 in HeLa cells expressing GFP-Sec22b treated with LLOMe or untreated (control). Arrowheads, colocalization. Scale bars, 5 μm. (I) GFP-Sec22b and LC3 overlapping area obtained by high content microscopy. (J) Co-IP analysis between flag-TRIM16 with GFP-Sec22b or GFP-syntaxin-17 co-expressed in HEK293T cells. (K) Co-IP analysis of endogenous proteins, TRIM16 and Sec22b, in lysates from THP-1 cells treated with LLOMe. (L) Co-IP analysis of interactions between deletion variants of flag-TRIM16 with GFP-Sec22b in HEK293T cells. (M and N) IL-1β secretion complementation experiments of TRIM16KO cells with TRIM16 Sec22b-interacting and its Sec22b-nonbinding mutant shown. (O) Two color super-resolution image showing co-clustering of flag-TRIM16 (red) and GFP-Sec22b (green); Scale bar, 200 nm. (P) Cross-correlation analysis of flag-TRIM16 and GFP-Sec22b super-resolution data showing a characteristic separation of ~ 70 nm.
Figure 1(A) Muscular exophers contain organelles and large protein complexes. Arrows: white - exopher, blue - muscle cell, green - mitochondria, red - proteasome foci. MOM - mitochondrial outer membrane. (B) BMW actively releases significant amounts of exopher. The image shows the middle part of the worm's body with muscles marked with dashed lines. Arrows indicate representative exophers, and the asterisk indicates the position of the vulva. (C) The ultrastructure of the muscular exopher and its schematic view. Arrows: red - morphologically changed mitochondria inside the exopher, yellow - normal, elongated mitochondria inside the muscle cell. c - cuticle, m - muscle, e - exopher. (D) Comparison between muscular exopher and coelomocyte. Arrows: white - exopher, blue - muscle, green- coelomocyte. (E) Exophers are formed via a pinching-off mechanism. Arrows: white - exopher, blue - muscle cell, red - distorted muscle cell membrane during exopher formation. (F) Production of muscular exophers is not evenly distributed across all muscle cells. The highest number of exophers is produced by the muscles adjacent to the vulva. n = 46; N = 3. (G) Exophers may remain connected to the sending BWM cells via thin elastic tubes that allow further transfer of cellular material. Arrows: white - exopher, blue - muscle cell, red - cellular material transferred to exopher via elastic tube. (H) Knockdown of two autophagy genes, atg-7 and lgg-1 significantly reduce the number of generated exophers. n = 91-103; N = 3. (I) Proteostasis disruption by hsf-1 knockdown does not increase exopher production. n = 60 and 55; N = 2. (J) Challenging proteostasis via oxidative stress but not heat stress increases exophers production. n = 30; N = 3.
Figure 5(A) HeLa cells were cultured in normal medium (NM) or glucose starvation (GS) for 4 h. The starved lysates were treated with calf intestinal phosphatase (CIP) to remove phosphate groups and analyzed by immunoblotting (IB) with Phos-tag gel or regular SDS-PAGE. (B) The cell lysates from HeLa cells were collected and analyzed after GS for indicated time (lane 1~5), or GS for 2 h followed by 2 h chase in NM (lane 6). The cell lysates were subjected to Phos-tag gel and regular SDS PAGE respectively. (C) WT or AMPKa1/2 double knockout MEFs were incubated under NM or GS for 4 h, followed by IB with Phos-tag gel or regular SDS-PAGE. (D) HeLa cells were incubated with compound C (20 mM, 30 min) with or without GS, followed by IB with Phos-tag gel or regular SDS-PAGE. (E) Bacterial purified His-tagged NH2-terminal 71 amino acid of PAQR3 was incubated with or without purified Flag-tagged ATG14L. Then the complexes were treated with WT AMPK or DN-AMPK (dominant negative AMPK) for the indicated time in vitro. PAQR3 phosphorylation was examined by Phos-tag gel. (F) HeLa cells infected with control or ATG14L-expressing lentivirus were incubated under GS for different time as indicated. Whole cell lysates were analyzed by both Phos-tag gel and regular SDS-PAGE. (G) HeLa cells were infected by lentivirus expressing control shRNA or ATG14L-specific shRNA. Then the ATG14L knockdown cells were stably re-expressed with shRNA-resistant ATG14L (rATG14L) or Beclin1 and incubated under NM or GS for 4 h. Whole cell lysates were analyzed by both Phos-tag gel and regular SDS-PAGE.
a–d) ELISA for IL-1β (a–c), IL-18 (d) and IL-6 (e) in supernatants of resting or LPS-primed bone marrow-derived macrophages (BMMs) stimulated with palmitate conjugated to BSA (PA-BSA) or BSA control.figf1(a-d) ELISA for IL-1β (a-c), IL-18 (d) and IL-6 (e) in supernatants of resting or LPS-primed bone marrow-derived macrophages (BMMs) stimulated with palmitate conjugated to BSA (PA-BSA) or BSA control. (e,f) ELISA for IL-1β (e) and IL-18 (f) in supernatants. Resting or LPS-primed BMMs generated from wild-type (WT), Nlrp3−/−, Pycard−/− or Nlrc4−/− mice were stimulated with PA-BSA. (g) IL-1β ELISA of supernatants from LPS-primed BMMs stimulated with PA-BSA in the absence or presence of the pan-caspase inhibitor zVAD or caspase-1 inhibitor zYVAD. (h) ELISA for IL-1β in supernatants. Resting or LPS-primed BMMs generated from WT or Casp1-/- mice were stimulated with PA-BSA, BSA control, ATP or nigericin. (i) ELISA for TNF in supernatants. Resting or LPS-primed BMMs generated from WT, Nlrp3-/-, Pycard-/- or Casp1-/- mice were stimulated with PA-BSA as indicated. Values are expressed as mean ± s.d., and the results are representative of three independent experiments.
Figure 4A. Schematic representation of OFD1 domain organization, with the predicted PKA phosphorylation sites and the position of S735 and S899 in the human variant. B. GST-fused, recombinant OFD1 polypeptides were used as substrates for in vitro phosphorylation assays using purified PKAc. GST-immobilized fusions were immunoblotted with an anti-phospho-(K/R)(K/R) X(S*/T*) antibody. A representative set of 3 independent experiments is shown. A Ponceau S staining of recombinant GST-OFD1 polypeptides is shown on the right panel. *Degradation products. C. HEK293 cells transiently expressing flag-OFD1 or flag-S735A mutant were starved for 24 hours and then treated with forskolin (FSK, 40μM/1 hour). Where indicated, cells were pre-treated with H89 (10μM). Lysates were subjected to flag immunoprecipitation and immunoblot analysis with anti-phospho-PKA substrates and anti-flag antibodies. Anti-GFP IgG were used as control. D. Quantitative analysis of the experiments shown in C. A mean value ± SD of three independent experiments is shown. Student's t test, *p<0,05, **p<0,01. E E. Same as in C, with the exception that cells were co-transfected with flag-OFD1 vector and with control siRNA (siCNT) or siRNA targeting praja2. F. Quantitative analysis of the experiments shown in E. A mean value ± SD of three independent experiments is shown. Student's t test, *p<0,05. G G. Schematic picture of the assembled TBC1D31 complex showing that following cAMP stimulation PKA phosphorylates the co-assembled OFD1.
Figure 2A. A pcDNA3.1 plasmid containing PTENα or canonical PTEN with a C-terminal FLAG tag was used for the detection of PTENα, PTENβ, canonical PTEN, and other unknown PTEN isoforms (upper panel); Indicated plasmids in the upper panel were transfected into HEK293 cells followed by western blotting analysis using FLAG antibody. GAPDH was used as a control (lower panel). B. The sequence of the 5′ UTR region of Homo sapiens PTEN mRNA. The codons that differ from AUG by only one nucleotide are highlighted in red. The initiation codons of PTENα, PTENβ, and canonical PTEN are separately highlighted in blue, yellow, and green. The potential initiation codons of PTENε are highlighted in red boxes. C. A different set of PTEN and PTENα constructs with a C-terminal FLAG tag, in which one of two sites (UCU783 or AGA942) was mutated to a UAG stop codon. D. Mutation of AGA942 into UAG eliminates PTENε expression, while the UCU783>UAG mutation has no such effect. C-terminal FLAG-tagged PTENα expression plasmids with and without UCU783>UAG or AGA942>UAG mutation were introduced into HEK293 cells separately, followed by immunoblotting with FLAG and GAPDH antibodies. E. A different set of PTENα constructs with a C-terminal FLAG tag, in which one of the non-AUG initiation codons between UCU783 and AGA942 (AGG810, CUG816, AAG831, CUG846, AAG936) was mutated to the CUC codon. F. Mutation of CUG816 but not any other potential non-AUG initiation codons eliminates PTENε expression. C-terminal FLAG-tagged PTENα expression plasmids with or without mutations were introduced into HEK293 cells separately, followed by immunoblotting with FLAG and GAPDH antibodies.
Figure 7A) Larval brains at 24 h ALH from wild-type control (UAS-β-Gal RNAi; UAS-β-Gal RNAi), msps RNAi control (UAS-GFP + UAS-msps RNAi (21982)), and rescued animals (UAS-klp64D+ msps RNAi (21982)) under the control of insc-Gal4 were analyzed for EdU incorporation, and larval brains were labeled with EdU and Dpn. B) Quantification graph of EdU-negative NSCs per brain lobe for genotypes in (A). n=13 BL for contorl; n=13 BL for msps RNAi; n=20 BL for UAS-klp64D + msps RNAi. ****p<0.0001. C) Quantification graph of NSC diameter for genotypes in (A). n=246 NSCs for control; n=494 NSCs for msps RNAi; n=457 NSCs for UAS-klp64D + msps RNAi. ****p<0.0001. D) Larval brains at 24 h ALH from wild-type control (UAS-β-Gal RNAi), RNAi control (UAS-msps RNAi (21982)), and rescued animals (g-kap3 + msps RNAi (21982)) driven by insc-Gal4 were examined for EdU incorporation, and larval brains were stained with EdU and Dpn. E) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (D). n=13 BL for control; n=15 BL for msps RNAi; n=19 BL for g-kap3 + msps RNAi. ****p<0.0001. F) Quantification graph of NSC diameter for genotypes in (D). n=339 NSCs for control; n=357 NSCs for msps RNAi; n=409 NSCs for g-kap3 + msps RNAi. Error bars represent SD.****p<0.0001. G) 24 h ALH Larval brains from wild-type control (UAS-β-Gal RNAi; UAS-β-Gal RNAi), RNAi control (UAS-GFP + UAS-msps RNAi (21982)), and rescued animals (UAS-E-cad7 + msps RNAi (21982)) were assessed for EdU incorporation, and larval brains were stained with EdU and Dpn. H) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (G). n=21 BL for control; n=21 BL for msps RNAi; n=22 BL for UAS-E-cad7 + msps RNAi. ****p<0.0001. I) Quantification graph of NSC diameter for genotypes in (G). n= n=127 NSCs for control; n=230 NSCs for msps RNAi; n=210 NSCs for UAS-E-cad7 + msps RNAi. ****p<0.0001. J) 24 h ALH Larval brains from wild-type control (UAS-β-Gal RNAi), mutant control (UAS-β-Gal RNAi + klp64Dk5h), and rescued animals (UAS-E-cad7 + klp64Dk5h) were processed for EdU incorporation, and larval brains were probed by with EdU, Dpn, and Mira. K) Quantification graph of NSCs negative for EdU incorporation per brain lobe for genotypes in (J). n=10 BL for control; n=12 BL for klp64Dk5h; n=10 BL for UAS-E-cad7 + klp64Dk5h. ****p<0.0001; p=0.3006 (ns). L) A working model. Microtubule arrays in the primary protrusion of Drosophila qNSCs are predominantly acentrosomal with plus-end-out orientation. Msps regulates microtubule growth and orientation in quiescent NSCs. Msps and Kinesin-2 target E-cadherin to the NSC-neuropil contact for NSC reactivation.