mbp  (Abcam)

 
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    5 5 Chlorosulfonyl 2 ethoxyphenyl 1 methyl 3 propyl 1 6 dihydro 7
    Description:

    Catalog Number:
    AB144950
    Price:
    None
    Buy from Supplier


    Structured Review

    Abcam mbp
    Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into <t>MBP</t> Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human <t>GFAP</t> (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p

    https://www.bioz.com/result/mbp/product/Abcam
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mbp - by Bioz Stars, 2021-07
    94/100 stars

    Images

    1) Product Images from "Familial t(1;11) translocation is associated with disruption of white matter structural integrity and oligodendrocyte-myelin dysfunction"

    Article Title: Familial t(1;11) translocation is associated with disruption of white matter structural integrity and oligodendrocyte-myelin dysfunction

    Journal: bioRxiv

    doi: 10.1101/657163

    Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into MBP Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human GFAP (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p
    Figure Legend Snippet: Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into MBP Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human GFAP (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p

    Techniques Used: Transplantation Assay

    Efficient conversion of control and case iPS cells to oligodendroglial lineage (a1-a7) Both control and case iPS lines could be patterned to OLIG2+ precursor cells under conditions described in Methods. (b1-b7) Further differentiation of cells from (a) gave PDFGRα+ OPCs under proliferative conditions containing FGF2 and PDGF-AA (c1-c7) Removal of mitogens produced mature oligodendrocytes that co-stained for O4 and MBP. (d-f) Quantification of OLIG2+ cells on day1 (d) PDFGRα+ OPCs on day7 (e) and O4+ oligodendrocytes at day21 (f) shows a significant increase in oligodendrocytes in case lines (n= 3 independent conversions for each line, unpaired t-test used to analyze p-values) Scale: (a-c) 50 μm
    Figure Legend Snippet: Efficient conversion of control and case iPS cells to oligodendroglial lineage (a1-a7) Both control and case iPS lines could be patterned to OLIG2+ precursor cells under conditions described in Methods. (b1-b7) Further differentiation of cells from (a) gave PDFGRα+ OPCs under proliferative conditions containing FGF2 and PDGF-AA (c1-c7) Removal of mitogens produced mature oligodendrocytes that co-stained for O4 and MBP. (d-f) Quantification of OLIG2+ cells on day1 (d) PDFGRα+ OPCs on day7 (e) and O4+ oligodendrocytes at day21 (f) shows a significant increase in oligodendrocytes in case lines (n= 3 independent conversions for each line, unpaired t-test used to analyze p-values) Scale: (a-c) 50 μm

    Techniques Used: Produced, Staining

    2) Product Images from "TLR4 response mediates ethanol-induced neurodevelopment alterations in a model of fetal alcohol spectrum disorders"

    Article Title: TLR4 response mediates ethanol-induced neurodevelopment alterations in a model of fetal alcohol spectrum disorders

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-0918-2

    Role of TLR4 in the expression of several proteins induced by prenatal and postnatal ethanol exposure. Data represent the immunoblot analysis of PLP, MBP, synaptotagmin, synapsin IIa, Tuj-1, CD11b, MHC-II, caspase 3 (active fragment of 17 kDa), and the ELISA analysis of MAP-2 from the cerebral cortices of the WT and TLR4-KO pups on PND 0, 20, and 66 exposed (E), or not (C), to ethanol during the embryonic and postnatal periods. A representative immunoblot of each protein is shown. PPEE prenatal and postnatal ethanol exposure. Data represent mean ± SEM, n = 8 mice/group. * p
    Figure Legend Snippet: Role of TLR4 in the expression of several proteins induced by prenatal and postnatal ethanol exposure. Data represent the immunoblot analysis of PLP, MBP, synaptotagmin, synapsin IIa, Tuj-1, CD11b, MHC-II, caspase 3 (active fragment of 17 kDa), and the ELISA analysis of MAP-2 from the cerebral cortices of the WT and TLR4-KO pups on PND 0, 20, and 66 exposed (E), or not (C), to ethanol during the embryonic and postnatal periods. A representative immunoblot of each protein is shown. PPEE prenatal and postnatal ethanol exposure. Data represent mean ± SEM, n = 8 mice/group. * p

    Techniques Used: Expressing, Plasmid Purification, Enzyme-linked Immunosorbent Assay, Mouse Assay

    Role of TLR4 in prenatal and postnatal ethanol exposure in PLP, MBP, and Iba-1 immunoreactivity in the cortices of the WT and TLR4-KO pups. Arrows show examples of microglial activation. The scale bar is 50 μm. Bars represent the quantification values of PLP, MBP, and Iba-1 immunoreactivity, expressed as the thresholded area occupied by specific staining in the cortices of the WT and TLR4-KO mice treated, or not, prenatally and postnatally with ethanol (PPEE). Results are given as means ± SEM ( n = 5). p
    Figure Legend Snippet: Role of TLR4 in prenatal and postnatal ethanol exposure in PLP, MBP, and Iba-1 immunoreactivity in the cortices of the WT and TLR4-KO pups. Arrows show examples of microglial activation. The scale bar is 50 μm. Bars represent the quantification values of PLP, MBP, and Iba-1 immunoreactivity, expressed as the thresholded area occupied by specific staining in the cortices of the WT and TLR4-KO mice treated, or not, prenatally and postnatally with ethanol (PPEE). Results are given as means ± SEM ( n = 5). p

    Techniques Used: Plasmid Purification, Activation Assay, Staining, Mouse Assay

    3) Product Images from "Oligodendrocyte differentiation from adult multipotent stem cells is modulated by glutamate"

    Article Title: Oligodendrocyte differentiation from adult multipotent stem cells is modulated by glutamate

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2011.144

    Myelination of cortical neurons by oligodendrocytes derived from SVZ neurospheres. ( a ) Primary cortical neurons stained with SMI31 at 3 and 8 DIV. Cell nuclei were counterstained with Hoechst 33258. Scale bar=20 μ m. ( b ) Scheme illustrating the temporal profile of individual and conjunct cultures. Only NG2-positive neurospheres were co-cultivated with primary neurons. Myelination in cocultures was assessed at 5 DIV. ( c and d ) Neurosphere sorting was assessed by immunofluorescence for NG2 (red) and Sox2 (green) before ( c ) and after ( d ) immunoselection. Yellow staining and arrowheads indicate NG2-positive neurospheres. Scale bar=100 μ m. ( e and g ) Representative fields showing the myelination of axons in control, untreated cultures ( e ) and cultures treated with 1 mM glutamate ( f ) or 100 μ M NMDA ( g ). ( h ) Histogram illustrating the degree of myelination as a percentage of axonal structures (NFL) co-localized with MBP (filled bars) or as myelin elongation expressed in micrometers (empty bars). All experiments with NMDA were performed in the presence of 100 μ M glycine. Data represent mean±S.E.M. of three independent experiments (10–12 fields each). * P
    Figure Legend Snippet: Myelination of cortical neurons by oligodendrocytes derived from SVZ neurospheres. ( a ) Primary cortical neurons stained with SMI31 at 3 and 8 DIV. Cell nuclei were counterstained with Hoechst 33258. Scale bar=20 μ m. ( b ) Scheme illustrating the temporal profile of individual and conjunct cultures. Only NG2-positive neurospheres were co-cultivated with primary neurons. Myelination in cocultures was assessed at 5 DIV. ( c and d ) Neurosphere sorting was assessed by immunofluorescence for NG2 (red) and Sox2 (green) before ( c ) and after ( d ) immunoselection. Yellow staining and arrowheads indicate NG2-positive neurospheres. Scale bar=100 μ m. ( e and g ) Representative fields showing the myelination of axons in control, untreated cultures ( e ) and cultures treated with 1 mM glutamate ( f ) or 100 μ M NMDA ( g ). ( h ) Histogram illustrating the degree of myelination as a percentage of axonal structures (NFL) co-localized with MBP (filled bars) or as myelin elongation expressed in micrometers (empty bars). All experiments with NMDA were performed in the presence of 100 μ M glycine. Data represent mean±S.E.M. of three independent experiments (10–12 fields each). * P

    Techniques Used: Derivative Assay, Staining, Immunofluorescence

    4) Product Images from "Directed Differentiation of Human Bone Marrow Stromal Cells to Fate-Committed Schwann Cells"

    Article Title: Directed Differentiation of Human Bone Marrow Stromal Cells to Fate-Committed Schwann Cells

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2017.08.004

    hBMSC-dSCs Myelinated Host Axons by Being Seeded into a Nerve Guide that Bridged a Critical Gap in a Rat Model of Sciatic Nerve Injury Longitudinal sections made in the mid-region of the sciatic nerve guide reveal the following. (A) Uni-axially aligned fibers immunopositive for rat TUJ1, representative of regrowing fibers. (B) Rows of Hoechst-stained nuclei between longitudinal layers immunopositive for human MBP. (C) Myelin-ensheathed axons and rows of peripherally located nuclei reminiscent of those of Schwann cells in the merged images of (A) and (B). (D) The myelin structure was illustrated in the TEM image of the transverse section (enlarged in d ∗ ). Scale bars, 50 μm for (A)–(C) and 200 nm for (D).
    Figure Legend Snippet: hBMSC-dSCs Myelinated Host Axons by Being Seeded into a Nerve Guide that Bridged a Critical Gap in a Rat Model of Sciatic Nerve Injury Longitudinal sections made in the mid-region of the sciatic nerve guide reveal the following. (A) Uni-axially aligned fibers immunopositive for rat TUJ1, representative of regrowing fibers. (B) Rows of Hoechst-stained nuclei between longitudinal layers immunopositive for human MBP. (C) Myelin-ensheathed axons and rows of peripherally located nuclei reminiscent of those of Schwann cells in the merged images of (A) and (B). (D) The myelin structure was illustrated in the TEM image of the transverse section (enlarged in d ∗ ). Scale bars, 50 μm for (A)–(C) and 200 nm for (D).

    Techniques Used: Staining, Transmission Electron Microscopy

    In Vitro Myelination of the DRG Neuritic Network by hBMSC-dSCs (A) Phase-contrast image showing hBMSC-dSCs (arrows) associated with neurons as early as 48 hr in co-culture with the neuritic network of purified DRG neurons in neuron maintenance medium (a). Immunofluorescence for S100 and TUJ1 in a parallel culture showing hBMSC-dSC (arrows) abutting on the neurites (b; right panels, zoom-in views of the boxed areas i–iii). Following 14 days of myelination induction, myelin-like segments (double-headed arrows) were formed by hBMSC-dSCs along the neuritic networks as shown by phase contrast (c) and immunofluorescence for MBP (d). Scale bar, 100 μm. (B) hBMSCs in parallel co-culture with DRG neurons (arrows) showed a fibroblast-like morphology (a) and failed to form MBP-positive segments along neurites (b). Scale bar, 100 μm. (C) SCLCs in parallel co-culture with the neuritic network of DRG neurons (arrows) reverted to the myofibroblast phenotype (a) and failed to form MBP-positive segments along neurites (b). Scale bar, 100 μm. (D) Histogram showing myelinated segment counts in ten fields for hBMSC-dSC versus hardly any for hBMSC ( ∗∗ p
    Figure Legend Snippet: In Vitro Myelination of the DRG Neuritic Network by hBMSC-dSCs (A) Phase-contrast image showing hBMSC-dSCs (arrows) associated with neurons as early as 48 hr in co-culture with the neuritic network of purified DRG neurons in neuron maintenance medium (a). Immunofluorescence for S100 and TUJ1 in a parallel culture showing hBMSC-dSC (arrows) abutting on the neurites (b; right panels, zoom-in views of the boxed areas i–iii). Following 14 days of myelination induction, myelin-like segments (double-headed arrows) were formed by hBMSC-dSCs along the neuritic networks as shown by phase contrast (c) and immunofluorescence for MBP (d). Scale bar, 100 μm. (B) hBMSCs in parallel co-culture with DRG neurons (arrows) showed a fibroblast-like morphology (a) and failed to form MBP-positive segments along neurites (b). Scale bar, 100 μm. (C) SCLCs in parallel co-culture with the neuritic network of DRG neurons (arrows) reverted to the myofibroblast phenotype (a) and failed to form MBP-positive segments along neurites (b). Scale bar, 100 μm. (D) Histogram showing myelinated segment counts in ten fields for hBMSC-dSC versus hardly any for hBMSC ( ∗∗ p

    Techniques Used: In Vitro, Co-Culture Assay, Purification, Immunofluorescence

    5) Product Images from "Fractalkine/CX3CR1 Contributes to Endometriosis-Induced Neuropathic Pain and Mechanical Hypersensitivity in Rats"

    Article Title: Fractalkine/CX3CR1 Contributes to Endometriosis-Induced Neuropathic Pain and Mechanical Hypersensitivity in Rats

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00495

    Expressions of fractalkine (FKN) and its receptor CX3CR1 in graft tissue. (A) Immunohistochemical staining for FKN and CX3CR1 in the sciatic nerve of graft tissue. (B) We analyzed FKN and CX3CR1 expression in the sciatic nerve of graft tissue by 1-way ANOVA. (C) Western blotting analysis showed protein levels of membrane-bound FKN, sFKN, and CX3CR1 in graft tissue. (D) Quantification of membrane-bound FKN, sFKN, and CX3CR1 bands in graft tissue, and analysis with 1-way ANOVA. (E) Immunofluorescence staining showed that FKN (red) was mainly expressed in macrophages labeled with Iba1 (green) in graft tissue. (F) The fluorescent images of FKN were statistically graphed for mean density, macrophage-positive cells, DAPI-positive cells, and co-expression in cells under 1-way ANOVA. (G) Immunofluorescent images show that CX3CR1 (red) was highly expressed in nerve fibers as indicated by PGP9.5 (green). The white arrows indicate the co-expression of CX3CR1 and PGP9.5 in cells. (H) Quantitative analysis of CX3CR1-positive cells, PGP9.5-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. (I) CX3CR1 was highly expressed on the myelin sheath when co-stained with MBP (green) using immunohistochemical staining. The white arrows indicate the co-expression of CX3CR1 and MBP in cells. (J) Quantitative analysis of CX3CR1-positive cells, MBP-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. N = 8 rats per group. ** p
    Figure Legend Snippet: Expressions of fractalkine (FKN) and its receptor CX3CR1 in graft tissue. (A) Immunohistochemical staining for FKN and CX3CR1 in the sciatic nerve of graft tissue. (B) We analyzed FKN and CX3CR1 expression in the sciatic nerve of graft tissue by 1-way ANOVA. (C) Western blotting analysis showed protein levels of membrane-bound FKN, sFKN, and CX3CR1 in graft tissue. (D) Quantification of membrane-bound FKN, sFKN, and CX3CR1 bands in graft tissue, and analysis with 1-way ANOVA. (E) Immunofluorescence staining showed that FKN (red) was mainly expressed in macrophages labeled with Iba1 (green) in graft tissue. (F) The fluorescent images of FKN were statistically graphed for mean density, macrophage-positive cells, DAPI-positive cells, and co-expression in cells under 1-way ANOVA. (G) Immunofluorescent images show that CX3CR1 (red) was highly expressed in nerve fibers as indicated by PGP9.5 (green). The white arrows indicate the co-expression of CX3CR1 and PGP9.5 in cells. (H) Quantitative analysis of CX3CR1-positive cells, PGP9.5-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. (I) CX3CR1 was highly expressed on the myelin sheath when co-stained with MBP (green) using immunohistochemical staining. The white arrows indicate the co-expression of CX3CR1 and MBP in cells. (J) Quantitative analysis of CX3CR1-positive cells, MBP-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. N = 8 rats per group. ** p

    Techniques Used: Immunohistochemistry, Staining, Expressing, Western Blot, Immunofluorescence, Labeling

    6) Product Images from "Myelination of Neuronal Cell Bodies when Myelin Supply Exceeds Axonal Demand"

    Article Title: Myelination of Neuronal Cell Bodies when Myelin Supply Exceeds Axonal Demand

    Journal: Current Biology

    doi: 10.1016/j.cub.2018.02.068

    Increasing Myelin Production Induces Cell Body Wrapping in the Zebrafish and Mouse Spinal Cord (A) 5 dpf spinal cord of control and transgenic zebrafish overexpressing constitutively active human Akt1 in oligodendrocytes, Tg(mbp:hAkt1DD), in double oligodendrocyte and myelin reporter line (arrowheads, wrapped cell bodies; asterisks, oligodendrocytes). (B) The number of wrapped cell bodies is significantly increased in Tg(mbp:hAkt1DD) (p = 0.001; n = 25 control and n = 14 Tg(mbp:hAkt1DD); t test). (C–F) Individual oligodendrocytes (C; asterisks note their cell bodies; note cytoplasmic TagRFPt expression in Tg(mbp:hAkt1DD) cell in inset) have a similar myelin sheath number and length at 5 dpf, quantified in (D) (p = 0.077; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test) and (E) (p = 0.412; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test), but Tg(mbp:hAkt1DD) animals have more myelinated cell bodies (arrowheads), quantified in (F) (p = 0.011; n = 21 control and n = 30 Tg(mbp:hAkt1DD); Mann-Whitney test). (C’) Another example of an individual oligodendrocyte in Tg(mbp:hAkt1DD) showing ectopic myelination of multiple cell bodies (arrowheads; asterisk denotes oligodendrocyte cell body). (G) Immunostaining of myelin basic protein (red, MBP) and NeuN (magenta) in the dorsal horn of the cervical spinal cord. Arrowhead indicates a wrapped NeuN+ neuronal cell body. (H) Optical sections representing 1-μm increments in the axial plane. Rightmost panel is a maximum intensity projection of MBP. (I) Frequency of ectopic wrapping events is slightly increased in the cervical spinal cord of P30 PTEN fl/fl ; CNP-Cre transgenic mice (p = 0.029; Mann-Whitney test; n = 4 animals per genotype; each animal quantified as the median of 6 pooled histological sections). All graphs display mean and SD, except for (F) and (I), which display median and interquartile range. The scale bars represent 10 μm (A), 5 μm (C and C’), 1 μm (inset C), 50 μm (G), and 20 μm (H).
    Figure Legend Snippet: Increasing Myelin Production Induces Cell Body Wrapping in the Zebrafish and Mouse Spinal Cord (A) 5 dpf spinal cord of control and transgenic zebrafish overexpressing constitutively active human Akt1 in oligodendrocytes, Tg(mbp:hAkt1DD), in double oligodendrocyte and myelin reporter line (arrowheads, wrapped cell bodies; asterisks, oligodendrocytes). (B) The number of wrapped cell bodies is significantly increased in Tg(mbp:hAkt1DD) (p = 0.001; n = 25 control and n = 14 Tg(mbp:hAkt1DD); t test). (C–F) Individual oligodendrocytes (C; asterisks note their cell bodies; note cytoplasmic TagRFPt expression in Tg(mbp:hAkt1DD) cell in inset) have a similar myelin sheath number and length at 5 dpf, quantified in (D) (p = 0.077; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test) and (E) (p = 0.412; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test), but Tg(mbp:hAkt1DD) animals have more myelinated cell bodies (arrowheads), quantified in (F) (p = 0.011; n = 21 control and n = 30 Tg(mbp:hAkt1DD); Mann-Whitney test). (C’) Another example of an individual oligodendrocyte in Tg(mbp:hAkt1DD) showing ectopic myelination of multiple cell bodies (arrowheads; asterisk denotes oligodendrocyte cell body). (G) Immunostaining of myelin basic protein (red, MBP) and NeuN (magenta) in the dorsal horn of the cervical spinal cord. Arrowhead indicates a wrapped NeuN+ neuronal cell body. (H) Optical sections representing 1-μm increments in the axial plane. Rightmost panel is a maximum intensity projection of MBP. (I) Frequency of ectopic wrapping events is slightly increased in the cervical spinal cord of P30 PTEN fl/fl ; CNP-Cre transgenic mice (p = 0.029; Mann-Whitney test; n = 4 animals per genotype; each animal quantified as the median of 6 pooled histological sections). All graphs display mean and SD, except for (F) and (I), which display median and interquartile range. The scale bars represent 10 μm (A), 5 μm (C and C’), 1 μm (inset C), 50 μm (G), and 20 μm (H).

    Techniques Used: Transgenic Assay, Expressing, MANN-WHITNEY, Immunostaining, Mouse Assay

    7) Product Images from "Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells"

    Article Title: Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells

    Journal: Nature Communications

    doi: 10.1038/s41467-018-06102-7

    Cellular profiling of spontaneous and purposely generated mutant oligodendrocyte alleles. a Diagram indicating that shiverer mice harbor a ~20-kilobase (kb) homozygous deletion encompassing exons 2–7 of the MBP gene. A Sanger sequencing trace shows the breakpoint of the shiverer deletion. b Diagram indicating the location of the two gRNAs designed to target MYRF . A Sanger sequencing trace shows the location of the homozygous deletion of exon 1. c Quantification of transcription factors Olig2, Nkx2.2, and Sox10 at passage 3 of the differentiation protocol. n = 3 shiverer cell lines; n = 3 replicate wells per cell line; > 179,500 cells scored per well. n = 3 MYRF KO and wild-type (WT) mESC replicate wells per cell line; > 700 cells scored per well. Data are represented as means ± SEM. d Fluorescent images of WT iPSC, shiverer , and MYRF KO OPCs expressing canonical OPC markers Olig2, Nkx2.2, and Sox10. Scale bar, 50 µm. e Cell surface immunostaining of the immature oligodendrocyte marker O4, after treatment with T3, of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. f Representative images of differentiated OPCs immunostained for mature oligodendrocyte markers MBP and PLP1, 72 h post treatment with T3 of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. g Representative images of OPC/DRG co-cultures stained for MBP and neurofilament (NF) at day 10 from WT iPSC, shiverer , and MYRF KO OPCs stained for PLP1 or MBP after being co-cultured for 10 days with NF+ embryonic rat DRGs. Scale bar, 50 µm
    Figure Legend Snippet: Cellular profiling of spontaneous and purposely generated mutant oligodendrocyte alleles. a Diagram indicating that shiverer mice harbor a ~20-kilobase (kb) homozygous deletion encompassing exons 2–7 of the MBP gene. A Sanger sequencing trace shows the breakpoint of the shiverer deletion. b Diagram indicating the location of the two gRNAs designed to target MYRF . A Sanger sequencing trace shows the location of the homozygous deletion of exon 1. c Quantification of transcription factors Olig2, Nkx2.2, and Sox10 at passage 3 of the differentiation protocol. n = 3 shiverer cell lines; n = 3 replicate wells per cell line; > 179,500 cells scored per well. n = 3 MYRF KO and wild-type (WT) mESC replicate wells per cell line; > 700 cells scored per well. Data are represented as means ± SEM. d Fluorescent images of WT iPSC, shiverer , and MYRF KO OPCs expressing canonical OPC markers Olig2, Nkx2.2, and Sox10. Scale bar, 50 µm. e Cell surface immunostaining of the immature oligodendrocyte marker O4, after treatment with T3, of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. f Representative images of differentiated OPCs immunostained for mature oligodendrocyte markers MBP and PLP1, 72 h post treatment with T3 of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. g Representative images of OPC/DRG co-cultures stained for MBP and neurofilament (NF) at day 10 from WT iPSC, shiverer , and MYRF KO OPCs stained for PLP1 or MBP after being co-cultured for 10 days with NF+ embryonic rat DRGs. Scale bar, 50 µm

    Techniques Used: Generated, Mutagenesis, Mouse Assay, Sequencing, Expressing, Immunostaining, Marker, Staining, Cell Culture

    8) Product Images from "Myelination of Neuronal Cell Bodies when Myelin Supply Exceeds Axonal Demand"

    Article Title: Myelination of Neuronal Cell Bodies when Myelin Supply Exceeds Axonal Demand

    Journal: Current Biology

    doi: 10.1016/j.cub.2018.02.068

    Increasing Myelin Production Induces Cell Body Wrapping in the Zebrafish and Mouse Spinal Cord (A) 5 dpf spinal cord of control and transgenic zebrafish overexpressing constitutively active human Akt1 in oligodendrocytes, Tg(mbp:hAkt1DD), in double oligodendrocyte and myelin reporter line (arrowheads, wrapped cell bodies; asterisks, oligodendrocytes). (B) The number of wrapped cell bodies is significantly increased in Tg(mbp:hAkt1DD) (p = 0.001; n = 25 control and n = 14 Tg(mbp:hAkt1DD); t test). (C–F) Individual oligodendrocytes (C; asterisks note their cell bodies; note cytoplasmic TagRFPt expression in Tg(mbp:hAkt1DD) cell in inset) have a similar myelin sheath number and length at 5 dpf, quantified in (D) (p = 0.077; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test) and (E) (p = 0.412; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test), but Tg(mbp:hAkt1DD) animals have more myelinated cell bodies (arrowheads), quantified in (F) (p = 0.011; n = 21 control and n = 30 Tg(mbp:hAkt1DD); Mann-Whitney test). (C’) Another example of an individual oligodendrocyte in Tg(mbp:hAkt1DD) showing ectopic myelination of multiple cell bodies (arrowheads; asterisk denotes oligodendrocyte cell body). (G) Immunostaining of myelin basic protein (red, MBP) and NeuN (magenta) in the dorsal horn of the cervical spinal cord. Arrowhead indicates a wrapped NeuN+ neuronal cell body. (H) Optical sections representing 1-μm increments in the axial plane. Rightmost panel is a maximum intensity projection of MBP. (I) Frequency of ectopic wrapping events is slightly increased in the cervical spinal cord of P30 PTEN fl/fl ; CNP-Cre transgenic mice (p = 0.029; Mann-Whitney test; n = 4 animals per genotype; each animal quantified as the median of 6 pooled histological sections). All graphs display mean and SD, except for (F) and (I), which display median and interquartile range. The scale bars represent 10 μm (A), 5 μm (C and C’), 1 μm (inset C), 50 μm (G), and 20 μm (H).
    Figure Legend Snippet: Increasing Myelin Production Induces Cell Body Wrapping in the Zebrafish and Mouse Spinal Cord (A) 5 dpf spinal cord of control and transgenic zebrafish overexpressing constitutively active human Akt1 in oligodendrocytes, Tg(mbp:hAkt1DD), in double oligodendrocyte and myelin reporter line (arrowheads, wrapped cell bodies; asterisks, oligodendrocytes). (B) The number of wrapped cell bodies is significantly increased in Tg(mbp:hAkt1DD) (p = 0.001; n = 25 control and n = 14 Tg(mbp:hAkt1DD); t test). (C–F) Individual oligodendrocytes (C; asterisks note their cell bodies; note cytoplasmic TagRFPt expression in Tg(mbp:hAkt1DD) cell in inset) have a similar myelin sheath number and length at 5 dpf, quantified in (D) (p = 0.077; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test) and (E) (p = 0.412; n = 16 control and n = 25 Tg(mbp:hAkt1DD); t test), but Tg(mbp:hAkt1DD) animals have more myelinated cell bodies (arrowheads), quantified in (F) (p = 0.011; n = 21 control and n = 30 Tg(mbp:hAkt1DD); Mann-Whitney test). (C’) Another example of an individual oligodendrocyte in Tg(mbp:hAkt1DD) showing ectopic myelination of multiple cell bodies (arrowheads; asterisk denotes oligodendrocyte cell body). (G) Immunostaining of myelin basic protein (red, MBP) and NeuN (magenta) in the dorsal horn of the cervical spinal cord. Arrowhead indicates a wrapped NeuN+ neuronal cell body. (H) Optical sections representing 1-μm increments in the axial plane. Rightmost panel is a maximum intensity projection of MBP. (I) Frequency of ectopic wrapping events is slightly increased in the cervical spinal cord of P30 PTEN fl/fl ; CNP-Cre transgenic mice (p = 0.029; Mann-Whitney test; n = 4 animals per genotype; each animal quantified as the median of 6 pooled histological sections). All graphs display mean and SD, except for (F) and (I), which display median and interquartile range. The scale bars represent 10 μm (A), 5 μm (C and C’), 1 μm (inset C), 50 μm (G), and 20 μm (H).

    Techniques Used: Transgenic Assay, Expressing, MANN-WHITNEY, Immunostaining, Mouse Assay

    9) Product Images from "Astrocytes phagocytose focal dystrophies from shortening myelin segments in the optic nerve of Xenopus laevis at metamorphosis"

    Article Title: Astrocytes phagocytose focal dystrophies from shortening myelin segments in the optic nerve of Xenopus laevis at metamorphosis

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1506486112

    ON metamorphic remodeling does not lead to degeneration of myelinated axons. ( A ) ON cross-sectional cryosections immunolabeled for acetylated tubulin (blue, axons), Blbp (green, astrocytes), and Mbp (red, oligodendrocyte myelin). (Scale bar: 50 μm.)
    Figure Legend Snippet: ON metamorphic remodeling does not lead to degeneration of myelinated axons. ( A ) ON cross-sectional cryosections immunolabeled for acetylated tubulin (blue, axons), Blbp (green, astrocytes), and Mbp (red, oligodendrocyte myelin). (Scale bar: 50 μm.)

    Techniques Used: Immunolabeling

    10) Product Images from "Oligodendrocytes Are Targets of HIV-1 Tat: NMDA and AMPA Receptor-Mediated Effects on Survival and Development"

    Article Title: Oligodendrocytes Are Targets of HIV-1 Tat: NMDA and AMPA Receptor-Mediated Effects on Survival and Development

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.4740-14.2015

    In vivo Tat expression leads to decreased MBP and MAG. Western blot analysis of tissue lysate from transgenic mice after 3 month Tat induction showed decreased levels of both MBP and MAG. Protein expression was normalized to glyceraldehyde 3-phosphate
    Figure Legend Snippet: In vivo Tat expression leads to decreased MBP and MAG. Western blot analysis of tissue lysate from transgenic mice after 3 month Tat induction showed decreased levels of both MBP and MAG. Protein expression was normalized to glyceraldehyde 3-phosphate

    Techniques Used: In Vivo, Expressing, Western Blot, Transgenic Assay, Mouse Assay

    11) Product Images from "c-Jun N-terminal kinase 1 (JNK1) modulates oligodendrocyte progenitor cell architecture, proliferation and myelination"

    Article Title: c-Jun N-terminal kinase 1 (JNK1) modulates oligodendrocyte progenitor cell architecture, proliferation and myelination

    Journal: Scientific Reports

    doi: 10.1038/s41598-021-86673-6

    Myelin alterations in JNK1 KO are not related to axonal abundance. ( A ) MBP + (red) and SMI31 neurofilaments (green) expression in the cortex and CC of P7, P15 and P90 WT vs JNK1KO mice. At P7 images illustrate the deep layers of the motor cortex, nearby cingulum bundle. Quantification of the percentage of MBP + ( B ), and SMI31 ( C ) pixels in the cortex and in CC of WT vs JNK1KO mice, and their ratio ( D) . ( E ) Western blots of P7, P15 and P30 WT and JNK1KO cortices and ( F ) corpora callosa. Full length blots are presented in Suppl. Fig. 5 ( A —cortices, B— corpora callosa). Scale bars: 100 μm. WT wild type, P postnatal day, A.U. arbitrary units, MBP Myelin Basic Protein, SMI31 neurofilaments, CNPase 2′,3′-Cyclic-Nucleotide 3′-phosphodiesterase, MOG Myelin Oligodendrocyte Glycoprotein. *P
    Figure Legend Snippet: Myelin alterations in JNK1 KO are not related to axonal abundance. ( A ) MBP + (red) and SMI31 neurofilaments (green) expression in the cortex and CC of P7, P15 and P90 WT vs JNK1KO mice. At P7 images illustrate the deep layers of the motor cortex, nearby cingulum bundle. Quantification of the percentage of MBP + ( B ), and SMI31 ( C ) pixels in the cortex and in CC of WT vs JNK1KO mice, and their ratio ( D) . ( E ) Western blots of P7, P15 and P30 WT and JNK1KO cortices and ( F ) corpora callosa. Full length blots are presented in Suppl. Fig. 5 ( A —cortices, B— corpora callosa). Scale bars: 100 μm. WT wild type, P postnatal day, A.U. arbitrary units, MBP Myelin Basic Protein, SMI31 neurofilaments, CNPase 2′,3′-Cyclic-Nucleotide 3′-phosphodiesterase, MOG Myelin Oligodendrocyte Glycoprotein. *P

    Techniques Used: Expressing, Mouse Assay, Western Blot

    12) Product Images from "Optogenetic stimulation promotes Schwann cell proliferation, differentiation, and myelination in vitro"

    Article Title: Optogenetic stimulation promotes Schwann cell proliferation, differentiation, and myelination in vitro

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-40173-w

    Effect of OS on SC differentiation and myelination in SC-motor neuron (MN) coculture. Transfected SC-MN co-cultures with and without OS were analyzed at DIV 7 and 14 by immunostaining antibodies against MBP (green), tubulin beta III (TuJ1, red), and DAPI (blue). ( a ) Representative confocal images of the SC-MN coculture are shown. Scale bar; 50 µm. ( b – d ) The levels of Krox20 or MBP were determined through western blot analysis. ( b ) Representative immunoblots and ( c , d ) quantification of Krox20 and MBP protein levels are shown. Note the substantially enhanced expression levels of Krox20 and MBP with OS at DIV 7 and 14, respectively. Protein levels were normalized against the level of ß-actin, which was used as a loading control. Graph shows means ± SEM from five independent experiments (unpaired two-tailed- t test with Welch’s correction). ** p = 7.78 × 10 −3 (Krox20); ** p = 1.27 × 10 −3 (MBP).
    Figure Legend Snippet: Effect of OS on SC differentiation and myelination in SC-motor neuron (MN) coculture. Transfected SC-MN co-cultures with and without OS were analyzed at DIV 7 and 14 by immunostaining antibodies against MBP (green), tubulin beta III (TuJ1, red), and DAPI (blue). ( a ) Representative confocal images of the SC-MN coculture are shown. Scale bar; 50 µm. ( b – d ) The levels of Krox20 or MBP were determined through western blot analysis. ( b ) Representative immunoblots and ( c , d ) quantification of Krox20 and MBP protein levels are shown. Note the substantially enhanced expression levels of Krox20 and MBP with OS at DIV 7 and 14, respectively. Protein levels were normalized against the level of ß-actin, which was used as a loading control. Graph shows means ± SEM from five independent experiments (unpaired two-tailed- t test with Welch’s correction). ** p = 7.78 × 10 −3 (Krox20); ** p = 1.27 × 10 −3 (MBP).

    Techniques Used: Transfection, Immunostaining, Western Blot, Expressing, Two Tailed Test

    Inhibition of MBP expression in optogenetic-mediated SCs with Ca 2+ blocker treatment. Transfected SC-MN cocultures were treated with mibefradil, U73122, Thaps. + dantrolene, or no addition at DIV 5, and the concentration of each reagent was maintained throughout the culture until fixation for analysis. The LED irradiator was applied to coculture samples at DIV 7, which were then stimulated for 3 days. ( a ) Representative confocal images of SC-MN coculture stained with MBP, TuJ1, and DAPI at DIV 10 and ( b ) quantification of MBP + -SCs are shown. MBP expression was substantially enhanced with OS and, interestingly, the level of MBP protein was inhibited by mibefradil, U73122, or Thaps. + dantrolene. Graph shows means ± SEM from three independent experiments (ANOVA and unpaired two-tailed- t test with Welch’s correction). Scale bar, 50 µm. ** p = 3.98 × 10 −3 (Transf.); ** p = 3.80 × 10 −3 (mibefradil); ** p = 7.02 × 10 −3 (U73122); ** p = 7.033 × 10 −3 (dantrolene); ** p = 7.02 × 10 −3 (U73122).
    Figure Legend Snippet: Inhibition of MBP expression in optogenetic-mediated SCs with Ca 2+ blocker treatment. Transfected SC-MN cocultures were treated with mibefradil, U73122, Thaps. + dantrolene, or no addition at DIV 5, and the concentration of each reagent was maintained throughout the culture until fixation for analysis. The LED irradiator was applied to coculture samples at DIV 7, which were then stimulated for 3 days. ( a ) Representative confocal images of SC-MN coculture stained with MBP, TuJ1, and DAPI at DIV 10 and ( b ) quantification of MBP + -SCs are shown. MBP expression was substantially enhanced with OS and, interestingly, the level of MBP protein was inhibited by mibefradil, U73122, or Thaps. + dantrolene. Graph shows means ± SEM from three independent experiments (ANOVA and unpaired two-tailed- t test with Welch’s correction). Scale bar, 50 µm. ** p = 3.98 × 10 −3 (Transf.); ** p = 3.80 × 10 −3 (mibefradil); ** p = 7.02 × 10 −3 (U73122); ** p = 7.033 × 10 −3 (dantrolene); ** p = 7.02 × 10 −3 (U73122).

    Techniques Used: Inhibition, Expressing, Transfection, Concentration Assay, Irradiation, Staining, Two Tailed Test

    13) Product Images from "Leukodystrophy resembling Vanishing White Matter Disease is recapitulated by brain-specific depletion of apoptosis regulator MCL-1"

    Article Title: Leukodystrophy resembling Vanishing White Matter Disease is recapitulated by brain-specific depletion of apoptosis regulator MCL-1

    Journal: bioRxiv

    doi: 10.1101/2020.12.02.408138

    Abnormal neuroimaging and myelination in Mcl-1 -deleted mice, with different rescue effects caused by co-deletion of Bak or Bax . ( A ) Representative MRIs show white matter hyperintensity in Mcl-1 cKO mice. ( B ) Total ventricular volume is abnormal in Mcl-1 cKO mice and increases from P7-P21, while the volume of the 4 th ventricle decreases significantly by P21. ( C,D ) Representative images and quantitative analysis of MBP IHC show that myelination is markedly decreased in Mcl-1 cKO mice, rescued in Mcl-1 cKO /Bak +/- mice, and incompletely rescued in Mcl-1/Bax dKO mice. (D) Graphs show MBP+ area/total area within individual sections, normalized to the mean value in the control mice. *, ** and *** denote p
    Figure Legend Snippet: Abnormal neuroimaging and myelination in Mcl-1 -deleted mice, with different rescue effects caused by co-deletion of Bak or Bax . ( A ) Representative MRIs show white matter hyperintensity in Mcl-1 cKO mice. ( B ) Total ventricular volume is abnormal in Mcl-1 cKO mice and increases from P7-P21, while the volume of the 4 th ventricle decreases significantly by P21. ( C,D ) Representative images and quantitative analysis of MBP IHC show that myelination is markedly decreased in Mcl-1 cKO mice, rescued in Mcl-1 cKO /Bak +/- mice, and incompletely rescued in Mcl-1/Bax dKO mice. (D) Graphs show MBP+ area/total area within individual sections, normalized to the mean value in the control mice. *, ** and *** denote p

    Techniques Used: Mouse Assay, Immunohistochemistry

    14) Product Images from "Metformin treatment after the hypoxia-ischemia attenuates brain injury in newborn rats"

    Article Title: Metformin treatment after the hypoxia-ischemia attenuates brain injury in newborn rats

    Journal: Oncotarget

    doi: 10.18632/oncotarget.20779

    Metformin treatment attenuated brain atrophy, enhanced axonal repairation and promoted remyelination after HI injury Rats were treated with metformin daily for 7 consecutive days, and the brains from each group were acquired at 7 d post HI injury. (A) General observation of rat brain from each group at 7 d after HI injury. Scale bar = 1 cm. (B-C) Representative images of immunohistochemical staining for MAP-2 (B) and MBP (C). Scale bar = 200 μm. (D) Representative western blots of MAP-2 and MBP. (E-F) Quantification of ipsilateral MAP-2 and MBP area loss. ** P
    Figure Legend Snippet: Metformin treatment attenuated brain atrophy, enhanced axonal repairation and promoted remyelination after HI injury Rats were treated with metformin daily for 7 consecutive days, and the brains from each group were acquired at 7 d post HI injury. (A) General observation of rat brain from each group at 7 d after HI injury. Scale bar = 1 cm. (B-C) Representative images of immunohistochemical staining for MAP-2 (B) and MBP (C). Scale bar = 200 μm. (D) Representative western blots of MAP-2 and MBP. (E-F) Quantification of ipsilateral MAP-2 and MBP area loss. ** P

    Techniques Used: Immunohistochemistry, Staining, Western Blot

    15) Product Images from "Ginseng Gintonin Attenuates Lead-Induced Rat Cerebellar Impairments during Gestation and Lactation"

    Article Title: Ginseng Gintonin Attenuates Lead-Induced Rat Cerebellar Impairments during Gestation and Lactation

    Journal: Biomolecules

    doi: 10.3390/biom10030385

    Effects of gintonin on Pb-induced cerebellar changes in various protein expression levels. Representative immunoblots for BDNF, Sirt1, MBP, GAD67, LPAR1, synaptophysin, NMDAR1, Bax, Bcl2, cleaved-caspase3, Nrf2, Mn-SOD, IL-1β, and TNFα ( A , B ) in the cerebellum of pups from control, Pb, and Pb plus gintonin (Pb + GT) groups at postnatal day 21 (PND21). Relative optical density (ROD) of each of immunoblot bands for BDNF ( p = 0.0010, one-way analysis of variance), Sirt1 ( p = 0.0020, one-way analysis of variance), MBP ( p
    Figure Legend Snippet: Effects of gintonin on Pb-induced cerebellar changes in various protein expression levels. Representative immunoblots for BDNF, Sirt1, MBP, GAD67, LPAR1, synaptophysin, NMDAR1, Bax, Bcl2, cleaved-caspase3, Nrf2, Mn-SOD, IL-1β, and TNFα ( A , B ) in the cerebellum of pups from control, Pb, and Pb plus gintonin (Pb + GT) groups at postnatal day 21 (PND21). Relative optical density (ROD) of each of immunoblot bands for BDNF ( p = 0.0010, one-way analysis of variance), Sirt1 ( p = 0.0020, one-way analysis of variance), MBP ( p

    Techniques Used: Expressing, Western Blot

    16) Product Images from "NogoA Neutralization Promotes Axonal Restoration After White Matter Injury In Subcortical Stroke"

    Article Title: NogoA Neutralization Promotes Axonal Restoration After White Matter Injury In Subcortical Stroke

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-09705-0

    In vitro study of anti-NogoA administration. ( A ) Schematic experimental protocol of the in vitro analysis. ( B ) Anti-NogoA antibody treatment decreased NogoA protein expression compared with Anti-IgG antibody. ( C ) Axonal growth was observed in PC12-differentiated cells after anti-NogoA antibody administration using phase-contrast microscopy. ( D ) Immunocytochemistry images of PC12 cells showed that anti-NogoA antibody treatment increased expression of GAP-43, NF, MAP-2 and MBP markers in PC12 cells compared with Anti-IgG antibody administration. Abbreviations: GAP-43, growth associated protein 43; NF, neurofilament; MAP-2, microtubule-associated protein 2; MBP, myelin basic protein.
    Figure Legend Snippet: In vitro study of anti-NogoA administration. ( A ) Schematic experimental protocol of the in vitro analysis. ( B ) Anti-NogoA antibody treatment decreased NogoA protein expression compared with Anti-IgG antibody. ( C ) Axonal growth was observed in PC12-differentiated cells after anti-NogoA antibody administration using phase-contrast microscopy. ( D ) Immunocytochemistry images of PC12 cells showed that anti-NogoA antibody treatment increased expression of GAP-43, NF, MAP-2 and MBP markers in PC12 cells compared with Anti-IgG antibody administration. Abbreviations: GAP-43, growth associated protein 43; NF, neurofilament; MAP-2, microtubule-associated protein 2; MBP, myelin basic protein.

    Techniques Used: In Vitro, Expressing, Microscopy, Immunocytochemistry

    17) Product Images from "Disruption of glial cell development by Zika virus contributes to severe microcephalic newborn mice"

    Article Title: Disruption of glial cell development by Zika virus contributes to severe microcephalic newborn mice

    Journal: Cell Discovery

    doi: 10.1038/s41421-018-0042-1

    Human mAb provides full protection against ZIKV infection associated damages in the developmental brain. Sections of neonatal brains infected or mock infected at E15.5 were inspected at P3. Antibody or PBS was injected about 4–6 h before virus injection ( a – d ). a Sections of cortices stained for ZIKV, S100β, and DAPI. Right panels: quantification of ZIKV and S100β positive cells. ZIKV: Ctrl n = 7/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 1.7 × 10 −10 ; Ctrl ZV-67 + ZIKV: P = 7.2 × 10 −14 ; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.1180; PBS + ZIKV ZK2B10 + ZIKV: P = 1.7 × 10 −10 . S100 β: Ctrl n = 7/5, PBS + ZIKV n = 6/4, ZV-67 + ZIKV n = 6/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 0.0059; Ctrl ZV-67 + ZIKV: P = 0.0144; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.6523; PBS + ZIKV ZK2B10 + ZIKV: P = 0293. b Sections were stained for ionized calcium-binding adapter molecule 1 (Iba-1) and DAPI. Enlarged representative images of Iba1 + cells are shown in the right panel. c Corpus callosum sections stained for Olig1 and DAPI. Right panel: quantification of Olig1 + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 0.0097; Ctrl ZV-67 + ZIKV: P = 0.0074; Ctrl ZK2B10 + ZIKV: P = 0.3048; PBS + ZIKV ZV-67 + ZIKV: P = 0.8436; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0206. d Corpus callosum sections were stained for CNPase. Right panels: quantification of CNPase + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 3.7 × 10 −5 ; Ctrl ZV-67 + ZIKV: P = 5.1 × 10 −5 ; Ctrl ZK2B10 + ZIKV: P = 0.9537; PBS + ZIKV ZV-67 + ZIKV: P = 0.9867; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0003. e Antibody or PBS was injected about 0.5 h before virus injection at P0. Slices from P9 brains were stained for MBP. Right panel: quantification of the relative intensity of MBP. Ctrl n = 8/3, PBS + ZIKV n = 8/3, ZV-67 + ZIKV n = 8/3, ZK2B10 + ZIKV n = 10/3. Ctrl PBS + ZIKV: P = 5.9 × 10 −8 ; Ctrl ZV-67 + ZIKV: P = 4.1 × 10 −7 ; Ctrl ZK2B10 + ZIKV: P = 0.0134; PBS + ZIKV ZV-67 + ZIKV: P = 0.0048; PBS + ZIKV ZK2B10 + ZIKV: P = 6.2 × 10 −5 . All data are mean ± SEM. ns: no significant, * P
    Figure Legend Snippet: Human mAb provides full protection against ZIKV infection associated damages in the developmental brain. Sections of neonatal brains infected or mock infected at E15.5 were inspected at P3. Antibody or PBS was injected about 4–6 h before virus injection ( a – d ). a Sections of cortices stained for ZIKV, S100β, and DAPI. Right panels: quantification of ZIKV and S100β positive cells. ZIKV: Ctrl n = 7/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 1.7 × 10 −10 ; Ctrl ZV-67 + ZIKV: P = 7.2 × 10 −14 ; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.1180; PBS + ZIKV ZK2B10 + ZIKV: P = 1.7 × 10 −10 . S100 β: Ctrl n = 7/5, PBS + ZIKV n = 6/4, ZV-67 + ZIKV n = 6/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 0.0059; Ctrl ZV-67 + ZIKV: P = 0.0144; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.6523; PBS + ZIKV ZK2B10 + ZIKV: P = 0293. b Sections were stained for ionized calcium-binding adapter molecule 1 (Iba-1) and DAPI. Enlarged representative images of Iba1 + cells are shown in the right panel. c Corpus callosum sections stained for Olig1 and DAPI. Right panel: quantification of Olig1 + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 0.0097; Ctrl ZV-67 + ZIKV: P = 0.0074; Ctrl ZK2B10 + ZIKV: P = 0.3048; PBS + ZIKV ZV-67 + ZIKV: P = 0.8436; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0206. d Corpus callosum sections were stained for CNPase. Right panels: quantification of CNPase + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 3.7 × 10 −5 ; Ctrl ZV-67 + ZIKV: P = 5.1 × 10 −5 ; Ctrl ZK2B10 + ZIKV: P = 0.9537; PBS + ZIKV ZV-67 + ZIKV: P = 0.9867; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0003. e Antibody or PBS was injected about 0.5 h before virus injection at P0. Slices from P9 brains were stained for MBP. Right panel: quantification of the relative intensity of MBP. Ctrl n = 8/3, PBS + ZIKV n = 8/3, ZV-67 + ZIKV n = 8/3, ZK2B10 + ZIKV n = 10/3. Ctrl PBS + ZIKV: P = 5.9 × 10 −8 ; Ctrl ZV-67 + ZIKV: P = 4.1 × 10 −7 ; Ctrl ZK2B10 + ZIKV: P = 0.0134; PBS + ZIKV ZV-67 + ZIKV: P = 0.0048; PBS + ZIKV ZK2B10 + ZIKV: P = 6.2 × 10 −5 . All data are mean ± SEM. ns: no significant, * P

    Techniques Used: Infection, Injection, Staining, Binding Assay

    Oligodendrocyte development is disrupted by ZIKV infection. Sections of neonatal brains were infected or mock infected at E15.5 and inspected at P0, P3, and P5. a Sagittal sections of corpus callosum stained for MBP and DAPI. b Sagittal sections of corpus callosum stained for 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP) and DAPI, and quantified for CNP + cells. P0: Ctrl n = 3/3, ZIKV n = 4/3; P3: Ctrl n = 10/4, ZIKV n = 12/4, P = 0.0001; P5: Ctrl n = 9/3, ZIKV n = 9/3, P = 3 × 10 −7 . c Upper panel: P5 brain staining for oligodendrocyte transcription factor 1 (Olig1), ZIKV, and DAPI. Lower panels: corpus callosum sections from P3 brain stained for Olig1, BrdU (labeled for 1 h), and Ki67, and quantified for Olig1 + BrdU + Ki67 + cells per total Ki67 + Olig1 + cells. n = 7/4, P = 0.002. d Corpus callosum sections from P3 brains were stained for Olig1, BrdU (labeled for 24 h), and Ki67. Right panels: quantification of cell cycle exit: BrdU + Olig1 + Ki67 − cells per total BrdU + Olig1 + cells. Ctrl: n = 11/3, ZIKV: n = 9/3, P = 0.003; quantification of Ki67 + Olig1 + cells per total Olig1 + cells. Ctrl: n = 9/3, ZIKV: n = 8/3, P = 0.0003. e Images of corpus callosum regions from P5 were co-stained for adenomatous polyposis coli clone CC1 and oligodendrocyte transcription factor 2 (Olig2). Right panel: quantification of CC1 and Olig2 double-positive cells per total Olig2 + cells. n = 8/3, P = 9.8 × 10 −8 . All data are mean ± SEM. ** P
    Figure Legend Snippet: Oligodendrocyte development is disrupted by ZIKV infection. Sections of neonatal brains were infected or mock infected at E15.5 and inspected at P0, P3, and P5. a Sagittal sections of corpus callosum stained for MBP and DAPI. b Sagittal sections of corpus callosum stained for 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP) and DAPI, and quantified for CNP + cells. P0: Ctrl n = 3/3, ZIKV n = 4/3; P3: Ctrl n = 10/4, ZIKV n = 12/4, P = 0.0001; P5: Ctrl n = 9/3, ZIKV n = 9/3, P = 3 × 10 −7 . c Upper panel: P5 brain staining for oligodendrocyte transcription factor 1 (Olig1), ZIKV, and DAPI. Lower panels: corpus callosum sections from P3 brain stained for Olig1, BrdU (labeled for 1 h), and Ki67, and quantified for Olig1 + BrdU + Ki67 + cells per total Ki67 + Olig1 + cells. n = 7/4, P = 0.002. d Corpus callosum sections from P3 brains were stained for Olig1, BrdU (labeled for 24 h), and Ki67. Right panels: quantification of cell cycle exit: BrdU + Olig1 + Ki67 − cells per total BrdU + Olig1 + cells. Ctrl: n = 11/3, ZIKV: n = 9/3, P = 0.003; quantification of Ki67 + Olig1 + cells per total Olig1 + cells. Ctrl: n = 9/3, ZIKV: n = 8/3, P = 0.0003. e Images of corpus callosum regions from P5 were co-stained for adenomatous polyposis coli clone CC1 and oligodendrocyte transcription factor 2 (Olig2). Right panel: quantification of CC1 and Olig2 double-positive cells per total Olig2 + cells. n = 8/3, P = 9.8 × 10 −8 . All data are mean ± SEM. ** P

    Techniques Used: Infection, Staining, Labeling

    18) Product Images from "Experience-Dependent Changes in Myelin Basic Protein Expression in Adult Visual and Somatosensory Cortex"

    Article Title: Experience-Dependent Changes in Myelin Basic Protein Expression in Adult Visual and Somatosensory Cortex

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2020.00056

    Expression of myelin basic protein (MBP) and Ube3A in S1 and V1. (A) S-EE increased MBP expression (+56%, SEM 30%, p
    Figure Legend Snippet: Expression of myelin basic protein (MBP) and Ube3A in S1 and V1. (A) S-EE increased MBP expression (+56%, SEM 30%, p

    Techniques Used: Expressing

    Hierarchical cluster analysis of adult V1. The high dimensional pattern of protein expression changes in V1 for the deprived (contralateral) and non-deprived (ipsilateral) hemispheres were analyzed by combining the measurements of MBP and Ube3A from this study with data for GluA2, PSD95, Gephyrin, Synapsin, and Synaptophysin from our previous study (Beshara et al., 2015 ). Correlation matrices are plotted to show the strength and direction (blue: negative; red: positive) of the pairwise Pearson’s R correlations between proteins for each condition and hemisphere. The inset with each panel shows the color-code and distribution of R values. The order of proteins was determined using unsupervised hierarchical clustering such that proteins with stronger correlations were nearby in the matrix: (A) normal, (B) fluoxetine, (C) MD contralateral (deprived) hemisphere, (D) fluoxetine and MD contralateral (deprived) hemisphere, (E) MD ipsilateral (non-deprived) hemisphere, and (F) fluoxetine and MD ipsilateral (non-deprived) hemisphere.
    Figure Legend Snippet: Hierarchical cluster analysis of adult V1. The high dimensional pattern of protein expression changes in V1 for the deprived (contralateral) and non-deprived (ipsilateral) hemispheres were analyzed by combining the measurements of MBP and Ube3A from this study with data for GluA2, PSD95, Gephyrin, Synapsin, and Synaptophysin from our previous study (Beshara et al., 2015 ). Correlation matrices are plotted to show the strength and direction (blue: negative; red: positive) of the pairwise Pearson’s R correlations between proteins for each condition and hemisphere. The inset with each panel shows the color-code and distribution of R values. The order of proteins was determined using unsupervised hierarchical clustering such that proteins with stronger correlations were nearby in the matrix: (A) normal, (B) fluoxetine, (C) MD contralateral (deprived) hemisphere, (D) fluoxetine and MD contralateral (deprived) hemisphere, (E) MD ipsilateral (non-deprived) hemisphere, and (F) fluoxetine and MD ipsilateral (non-deprived) hemisphere.

    Techniques Used: Expressing

    19) Product Images from "Luoyutong Treatment Promotes Functional Recovery and Neuronal Plasticity after Cerebral Ischemia-Reperfusion Injury in Rats"

    Article Title: Luoyutong Treatment Promotes Functional Recovery and Neuronal Plasticity after Cerebral Ischemia-Reperfusion Injury in Rats

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2015/369021

    Schematic representation of the experimental procedures. (Luoyutong: LYT; middle cerebral artery occlusion: MCAO; 2,3,5-triphenyltetrazolium chloride: TTC; microtubule associated protein: MAP-2; myelin basic protein: MBP; brain derived neurotrophic factor: BDNF; basic fibroblast growth factor: b-FGF; neuron-specific nuclear protein: NeuN; neurofilament 200: NF200).
    Figure Legend Snippet: Schematic representation of the experimental procedures. (Luoyutong: LYT; middle cerebral artery occlusion: MCAO; 2,3,5-triphenyltetrazolium chloride: TTC; microtubule associated protein: MAP-2; myelin basic protein: MBP; brain derived neurotrophic factor: BDNF; basic fibroblast growth factor: b-FGF; neuron-specific nuclear protein: NeuN; neurofilament 200: NF200).

    Techniques Used: Derivative Assay

    20) Product Images from "let-7 miRNAs Can Act through Notch to Regulate Human Gliogenesis"

    Article Title: let-7 miRNAs Can Act through Notch to Regulate Human Gliogenesis

    Journal: Stem Cell Reports

    doi: 10.1016/j.stemcr.2014.08.015

    Generation of Oligodendrocytes Is Facilitated by Induction of let-7 (A) At 6 weeks of differentiation toward the oligodendrocyte lineage, OPC markers were identified by immunostaining and quantified in (B). (B) Quantification of results presented in (A). (C) Following at least 6 weeks of differentiation in either base or glial induction medium, mature oligodendrocytes were identified by immunostaining and quantified in (D). The bottom panels followed a glial induction protocol ( Wang et al., 2013 ) and produced oligodendrocytes after just 6.5 weeks, again, only with let-7 induction at the NPC stage prior to directed differentiation. At 7.5 weeks, the number of oligodendrocytes produced was considerably larger. Scale bar, 100 mm. (D) A quantification of oligodendrocytes in each condition at two time points in each well. Cells were judged as bona fide oligodendrocytes if positive for both O4 and MBP.
    Figure Legend Snippet: Generation of Oligodendrocytes Is Facilitated by Induction of let-7 (A) At 6 weeks of differentiation toward the oligodendrocyte lineage, OPC markers were identified by immunostaining and quantified in (B). (B) Quantification of results presented in (A). (C) Following at least 6 weeks of differentiation in either base or glial induction medium, mature oligodendrocytes were identified by immunostaining and quantified in (D). The bottom panels followed a glial induction protocol ( Wang et al., 2013 ) and produced oligodendrocytes after just 6.5 weeks, again, only with let-7 induction at the NPC stage prior to directed differentiation. At 7.5 weeks, the number of oligodendrocytes produced was considerably larger. Scale bar, 100 mm. (D) A quantification of oligodendrocytes in each condition at two time points in each well. Cells were judged as bona fide oligodendrocytes if positive for both O4 and MBP.

    Techniques Used: Immunostaining, Produced

    21) Product Images from "MicroRNA-146a promotes oligodendrogenesis in stroke"

    Article Title: MicroRNA-146a promotes oligodendrogenesis in stroke

    Journal: Molecular neurobiology

    doi: 10.1007/s12035-015-9655-7

    miR-146a regulates oligodendrocyte differentiation by targeting IRAK1. Using a miR-146a target PCR array, panel A showed that the transcripts of IRAK1, IRAK2, TRAF6 and Numb were decreased in OPCs after overexpression of miR-146a by introducing exogenous miR-146a mimics, compared with those transfected with miRNA mimic control. Western blot demonstrated that overexpression of miR-146a decreased its target genes-IRAK1 and another target gene TRAF6 as well as NF-κβ p65 subunit in neural progenitor cells isolated from both contralateral and ischemic SVZ cells with a dose-dependent manner (B). Panel C shows that nanoparticle-delivered mature miR-146a resulted in a substantial decrease of IRAK1 protein levels compared with the miRNA mimics control as well as attenuation of endogenous miR-146a increased IRAK1 levels in primary cultured OPCs. Panel D shows that protein levels of IRAK1 were decreased in OPCs transfected with its siRNA and knockdown of endogenous IRAK1 increased protein levels of MBP, CNPase, PLP and cleaved Caspase-3 whereas decreased protein levels of NG2 and PDGFRa. Panel E shows that knockdown of IRAK1 significantly increased the Caspase-3/7 activity in OPCs.
    Figure Legend Snippet: miR-146a regulates oligodendrocyte differentiation by targeting IRAK1. Using a miR-146a target PCR array, panel A showed that the transcripts of IRAK1, IRAK2, TRAF6 and Numb were decreased in OPCs after overexpression of miR-146a by introducing exogenous miR-146a mimics, compared with those transfected with miRNA mimic control. Western blot demonstrated that overexpression of miR-146a decreased its target genes-IRAK1 and another target gene TRAF6 as well as NF-κβ p65 subunit in neural progenitor cells isolated from both contralateral and ischemic SVZ cells with a dose-dependent manner (B). Panel C shows that nanoparticle-delivered mature miR-146a resulted in a substantial decrease of IRAK1 protein levels compared with the miRNA mimics control as well as attenuation of endogenous miR-146a increased IRAK1 levels in primary cultured OPCs. Panel D shows that protein levels of IRAK1 were decreased in OPCs transfected with its siRNA and knockdown of endogenous IRAK1 increased protein levels of MBP, CNPase, PLP and cleaved Caspase-3 whereas decreased protein levels of NG2 and PDGFRa. Panel E shows that knockdown of IRAK1 significantly increased the Caspase-3/7 activity in OPCs.

    Techniques Used: Polymerase Chain Reaction, Over Expression, Transfection, Western Blot, Isolation, Cell Culture, Plasmid Purification, Activity Assay

    The effects of miR-146a on the differentiation and survival of oligodendrocyte progenitor cells (OPCs). Panels A and B demonstrate the introduction of miR-146a mimics (A) or inhibitors (B) significantly increased or decreased the expression of miR-146a in OPCs, respectively. Panel C shows representative immunostaining images of MBP positive cells after miR-146a mimic transfection. Panel D shows quantitative data of the number of MBP positive cells in OPCs after treatment with miR-146a mimics or inhibitors. OPCs transfected with cel-miR-67 mimics or inhibitors was employed as a negative control (D, control). Western blots (E) show that delivery of miR-146a mimics increased protein levels of MBP, proteolipid protein (PLP), and 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), markers of mature oligodendrocytes as well as considerably decreased oligodendrocyte progenitor cell protein levels, PDGFRa and NG2, nevertheless inhibition of miR-146a using inhibitor against miR-146a. Panel F shows representative BrdU positive cells in OPCs after transfection of miR-146a mimic. Panel G shows quantitative data of the number of BrdU positive cells in OPCs after treatment with miR-146a mimics. Panel H shows that delivery of miR-146a mimics dramatically decreased the Caspase-3/7 activity tested by a luciferase reporter in OPCs, but miR-146a inhibitor inversely increased the Caspase-3/7 activity. *p
    Figure Legend Snippet: The effects of miR-146a on the differentiation and survival of oligodendrocyte progenitor cells (OPCs). Panels A and B demonstrate the introduction of miR-146a mimics (A) or inhibitors (B) significantly increased or decreased the expression of miR-146a in OPCs, respectively. Panel C shows representative immunostaining images of MBP positive cells after miR-146a mimic transfection. Panel D shows quantitative data of the number of MBP positive cells in OPCs after treatment with miR-146a mimics or inhibitors. OPCs transfected with cel-miR-67 mimics or inhibitors was employed as a negative control (D, control). Western blots (E) show that delivery of miR-146a mimics increased protein levels of MBP, proteolipid protein (PLP), and 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), markers of mature oligodendrocytes as well as considerably decreased oligodendrocyte progenitor cell protein levels, PDGFRa and NG2, nevertheless inhibition of miR-146a using inhibitor against miR-146a. Panel F shows representative BrdU positive cells in OPCs after transfection of miR-146a mimic. Panel G shows quantitative data of the number of BrdU positive cells in OPCs after treatment with miR-146a mimics. Panel H shows that delivery of miR-146a mimics dramatically decreased the Caspase-3/7 activity tested by a luciferase reporter in OPCs, but miR-146a inhibitor inversely increased the Caspase-3/7 activity. *p

    Techniques Used: Expressing, Immunostaining, Transfection, Negative Control, Western Blot, Plasmid Purification, Inhibition, Activity Assay, Luciferase

    22) Product Images from "Disruption of glial cell development by Zika virus contributes to severe microcephalic newborn mice"

    Article Title: Disruption of glial cell development by Zika virus contributes to severe microcephalic newborn mice

    Journal: Cell Discovery

    doi: 10.1038/s41421-018-0042-1

    Human mAb provides full protection against ZIKV infection associated damages in the developmental brain. Sections of neonatal brains infected or mock infected at E15.5 were inspected at P3. Antibody or PBS was injected about 4–6 h before virus injection ( a – d ). a Sections of cortices stained for ZIKV, S100β, and DAPI. Right panels: quantification of ZIKV and S100β positive cells. ZIKV: Ctrl n = 7/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 1.7 × 10 −10 ; Ctrl ZV-67 + ZIKV: P = 7.2 × 10 −14 ; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.1180; PBS + ZIKV ZK2B10 + ZIKV: P = 1.7 × 10 −10 . S100 β: Ctrl n = 7/5, PBS + ZIKV n = 6/4, ZV-67 + ZIKV n = 6/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 0.0059; Ctrl ZV-67 + ZIKV: P = 0.0144; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.6523; PBS + ZIKV ZK2B10 + ZIKV: P = 0293. b Sections were stained for ionized calcium-binding adapter molecule 1 (Iba-1) and DAPI. Enlarged representative images of Iba1 + cells are shown in the right panel. c Corpus callosum sections stained for Olig1 and DAPI. Right panel: quantification of Olig1 + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 0.0097; Ctrl ZV-67 + ZIKV: P = 0.0074; Ctrl ZK2B10 + ZIKV: P = 0.3048; PBS + ZIKV ZV-67 + ZIKV: P = 0.8436; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0206. d Corpus callosum sections were stained for CNPase. Right panels: quantification of CNPase + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 3.7 × 10 −5 ; Ctrl ZV-67 + ZIKV: P = 5.1 × 10 −5 ; Ctrl ZK2B10 + ZIKV: P = 0.9537; PBS + ZIKV ZV-67 + ZIKV: P = 0.9867; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0003. e Antibody or PBS was injected about 0.5 h before virus injection at P0. Slices from P9 brains were stained for MBP. Right panel: quantification of the relative intensity of MBP. Ctrl n = 8/3, PBS + ZIKV n = 8/3, ZV-67 + ZIKV n = 8/3, ZK2B10 + ZIKV n = 10/3. Ctrl PBS + ZIKV: P = 5.9 × 10 −8 ; Ctrl ZV-67 + ZIKV: P = 4.1 × 10 −7 ; Ctrl ZK2B10 + ZIKV: P = 0.0134; PBS + ZIKV ZV-67 + ZIKV: P = 0.0048; PBS + ZIKV ZK2B10 + ZIKV: P = 6.2 × 10 −5 . All data are mean ± SEM. ns: no significant, * P
    Figure Legend Snippet: Human mAb provides full protection against ZIKV infection associated damages in the developmental brain. Sections of neonatal brains infected or mock infected at E15.5 were inspected at P3. Antibody or PBS was injected about 4–6 h before virus injection ( a – d ). a Sections of cortices stained for ZIKV, S100β, and DAPI. Right panels: quantification of ZIKV and S100β positive cells. ZIKV: Ctrl n = 7/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 1.7 × 10 −10 ; Ctrl ZV-67 + ZIKV: P = 7.2 × 10 −14 ; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.1180; PBS + ZIKV ZK2B10 + ZIKV: P = 1.7 × 10 −10 . S100 β: Ctrl n = 7/5, PBS + ZIKV n = 6/4, ZV-67 + ZIKV n = 6/4, ZK2B10 + ZIKV n = 7/4. Ctrl PBS + ZIKV: P = 0.0059; Ctrl ZV-67 + ZIKV: P = 0.0144; Ctrl ZK2B10 + ZIKV: P = 0.2221; PBS + ZIKV ZV-67 + ZIKV: P = 0.6523; PBS + ZIKV ZK2B10 + ZIKV: P = 0293. b Sections were stained for ionized calcium-binding adapter molecule 1 (Iba-1) and DAPI. Enlarged representative images of Iba1 + cells are shown in the right panel. c Corpus callosum sections stained for Olig1 and DAPI. Right panel: quantification of Olig1 + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 0.0097; Ctrl ZV-67 + ZIKV: P = 0.0074; Ctrl ZK2B10 + ZIKV: P = 0.3048; PBS + ZIKV ZV-67 + ZIKV: P = 0.8436; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0206. d Corpus callosum sections were stained for CNPase. Right panels: quantification of CNPase + cells. Ctrl n = 9/4, PBS + ZIKV n = 9/4, ZV-67 + ZIKV n = 9/4, ZK2B10 n = 9/4. Ctrl PBS + ZIKV: P = 3.7 × 10 −5 ; Ctrl ZV-67 + ZIKV: P = 5.1 × 10 −5 ; Ctrl ZK2B10 + ZIKV: P = 0.9537; PBS + ZIKV ZV-67 + ZIKV: P = 0.9867; PBS + ZIKV ZK2B10 + ZIKV: P = 0.0003. e Antibody or PBS was injected about 0.5 h before virus injection at P0. Slices from P9 brains were stained for MBP. Right panel: quantification of the relative intensity of MBP. Ctrl n = 8/3, PBS + ZIKV n = 8/3, ZV-67 + ZIKV n = 8/3, ZK2B10 + ZIKV n = 10/3. Ctrl PBS + ZIKV: P = 5.9 × 10 −8 ; Ctrl ZV-67 + ZIKV: P = 4.1 × 10 −7 ; Ctrl ZK2B10 + ZIKV: P = 0.0134; PBS + ZIKV ZV-67 + ZIKV: P = 0.0048; PBS + ZIKV ZK2B10 + ZIKV: P = 6.2 × 10 −5 . All data are mean ± SEM. ns: no significant, * P

    Techniques Used: Infection, Injection, Staining, Binding Assay

    Oligodendrocyte development is disrupted by ZIKV infection. Sections of neonatal brains were infected or mock infected at E15.5 and inspected at P0, P3, and P5. a Sagittal sections of corpus callosum stained for MBP and DAPI. b Sagittal sections of corpus callosum stained for 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP) and DAPI, and quantified for CNP + cells. P0: Ctrl n = 3/3, ZIKV n = 4/3; P3: Ctrl n = 10/4, ZIKV n = 12/4, P = 0.0001; P5: Ctrl n = 9/3, ZIKV n = 9/3, P = 3 × 10 −7 . c Upper panel: P5 brain staining for oligodendrocyte transcription factor 1 (Olig1), ZIKV, and DAPI. Lower panels: corpus callosum sections from P3 brain stained for Olig1, BrdU (labeled for 1 h), and Ki67, and quantified for Olig1 + BrdU + Ki67 + cells per total Ki67 + Olig1 + cells. n = 7/4, P = 0.002. d Corpus callosum sections from P3 brains were stained for Olig1, BrdU (labeled for 24 h), and Ki67. Right panels: quantification of cell cycle exit: BrdU + Olig1 + Ki67 − cells per total BrdU + Olig1 + cells. Ctrl: n = 11/3, ZIKV: n = 9/3, P = 0.003; quantification of Ki67 + Olig1 + cells per total Olig1 + cells. Ctrl: n = 9/3, ZIKV: n = 8/3, P = 0.0003. e Images of corpus callosum regions from P5 were co-stained for adenomatous polyposis coli clone CC1 and oligodendrocyte transcription factor 2 (Olig2). Right panel: quantification of CC1 and Olig2 double-positive cells per total Olig2 + cells. n = 8/3, P = 9.8 × 10 −8 . All data are mean ± SEM. ** P
    Figure Legend Snippet: Oligodendrocyte development is disrupted by ZIKV infection. Sections of neonatal brains were infected or mock infected at E15.5 and inspected at P0, P3, and P5. a Sagittal sections of corpus callosum stained for MBP and DAPI. b Sagittal sections of corpus callosum stained for 2′,3′-cyclic-nucleotide 3′-phosphodiesterase (CNP) and DAPI, and quantified for CNP + cells. P0: Ctrl n = 3/3, ZIKV n = 4/3; P3: Ctrl n = 10/4, ZIKV n = 12/4, P = 0.0001; P5: Ctrl n = 9/3, ZIKV n = 9/3, P = 3 × 10 −7 . c Upper panel: P5 brain staining for oligodendrocyte transcription factor 1 (Olig1), ZIKV, and DAPI. Lower panels: corpus callosum sections from P3 brain stained for Olig1, BrdU (labeled for 1 h), and Ki67, and quantified for Olig1 + BrdU + Ki67 + cells per total Ki67 + Olig1 + cells. n = 7/4, P = 0.002. d Corpus callosum sections from P3 brains were stained for Olig1, BrdU (labeled for 24 h), and Ki67. Right panels: quantification of cell cycle exit: BrdU + Olig1 + Ki67 − cells per total BrdU + Olig1 + cells. Ctrl: n = 11/3, ZIKV: n = 9/3, P = 0.003; quantification of Ki67 + Olig1 + cells per total Olig1 + cells. Ctrl: n = 9/3, ZIKV: n = 8/3, P = 0.0003. e Images of corpus callosum regions from P5 were co-stained for adenomatous polyposis coli clone CC1 and oligodendrocyte transcription factor 2 (Olig2). Right panel: quantification of CC1 and Olig2 double-positive cells per total Olig2 + cells. n = 8/3, P = 9.8 × 10 −8 . All data are mean ± SEM. ** P

    Techniques Used: Infection, Staining, Labeling

    23) Product Images from "Suppression of proteolipid protein rescues Pelizaeus-Merzbacher disease"

    Article Title: Suppression of proteolipid protein rescues Pelizaeus-Merzbacher disease

    Journal: Nature

    doi: 10.1038/s41586-020-2494-3

    Postnatal delivery of Plp1 -targeted antisense oligonucleotides rescues lifespan and oligodendrocytes with partial restoration of myelin in jimpy . a , Schematic of ASO experimental design used in Figures 3- 4 . b , Kaplan-Meier plot depicting the lifespan of contemporaneous wtASOctr, uninjected jimpy , jpASOctr, jpASO Plp1 .a, and jpASO Plp1 .b (n=12, 14, 5, 5 mice, respectively). p-values calculated using the log-rank test. See Supplementary Data 6 for source metadata. c-d , Immunohistochemical images of ( c ) 3-week and ( d ) 8-month whole-brain sagittal sections showing MBP + myelin (green) and DAPI + nuclei (blue) staining. Scale bar, 2mm. See Supplementary Data 8 – 10 for higher magnification. e-f , Quantification of ( e ) MyRF + oligodendrocytes and ( f ) Sox10 + glial lineage cells at 3 weeks of age (n=3 mice). For representative source images, see Supplementary Data 8 – 10 . g-h , ( g ) Electron micrograph images and ( h ) quantification of myelinated axons at 3 weeks of age (n=3–5 mice). Scale bar, 0.5μm. Biological replicates (individual mice) indicated by open circles. Graph bars indicate mean ± standard deviation. p-values calculated using one-way ANOVA with Dunnett’s correction, except where indicated. p-values stated for p
    Figure Legend Snippet: Postnatal delivery of Plp1 -targeted antisense oligonucleotides rescues lifespan and oligodendrocytes with partial restoration of myelin in jimpy . a , Schematic of ASO experimental design used in Figures 3- 4 . b , Kaplan-Meier plot depicting the lifespan of contemporaneous wtASOctr, uninjected jimpy , jpASOctr, jpASO Plp1 .a, and jpASO Plp1 .b (n=12, 14, 5, 5 mice, respectively). p-values calculated using the log-rank test. See Supplementary Data 6 for source metadata. c-d , Immunohistochemical images of ( c ) 3-week and ( d ) 8-month whole-brain sagittal sections showing MBP + myelin (green) and DAPI + nuclei (blue) staining. Scale bar, 2mm. See Supplementary Data 8 – 10 for higher magnification. e-f , Quantification of ( e ) MyRF + oligodendrocytes and ( f ) Sox10 + glial lineage cells at 3 weeks of age (n=3 mice). For representative source images, see Supplementary Data 8 – 10 . g-h , ( g ) Electron micrograph images and ( h ) quantification of myelinated axons at 3 weeks of age (n=3–5 mice). Scale bar, 0.5μm. Biological replicates (individual mice) indicated by open circles. Graph bars indicate mean ± standard deviation. p-values calculated using one-way ANOVA with Dunnett’s correction, except where indicated. p-values stated for p

    Techniques Used: Allele-specific Oligonucleotide, Mouse Assay, Immunohistochemistry, Staining, Standard Deviation

    Plp1 suppression in jimpy OPCs rescues survival of differentiating oligodendrocytes in vitro . a , Phase and immunocytochemistry images of Oct4 + and Nanog + iPSCs, along with DAPI + nuclei and b , normal karyotype of a CR- impy iPSC line used to generate OPCs. Scale bar, 50μm. c , Immunocytochemistry images showing Olig2 + and Sox10 + cells in OPC cultures, along with DAPI + nuclei, derived from iPSCs. Scale bar, 100μm. d , Percentage of Sox10 + and Olig2 + cells in OPC cultures. e , Immunocytochemistry images of MBP + and PLP + oligodendrocytes. f-g , Quantification of ( f ) MBP + oligodendrocytes and ( g ) total cell number (DAPI + nuclei) from iPSC-derived OPCs differentiated in vitro for 3 days. Scale bar, 50μm. Technical replicates (individual wells) for a single cell line per genotype indicated by black circles. Graph bars indicate mean ± standard deviation.
    Figure Legend Snippet: Plp1 suppression in jimpy OPCs rescues survival of differentiating oligodendrocytes in vitro . a , Phase and immunocytochemistry images of Oct4 + and Nanog + iPSCs, along with DAPI + nuclei and b , normal karyotype of a CR- impy iPSC line used to generate OPCs. Scale bar, 50μm. c , Immunocytochemistry images showing Olig2 + and Sox10 + cells in OPC cultures, along with DAPI + nuclei, derived from iPSCs. Scale bar, 100μm. d , Percentage of Sox10 + and Olig2 + cells in OPC cultures. e , Immunocytochemistry images of MBP + and PLP + oligodendrocytes. f-g , Quantification of ( f ) MBP + oligodendrocytes and ( g ) total cell number (DAPI + nuclei) from iPSC-derived OPCs differentiated in vitro for 3 days. Scale bar, 50μm. Technical replicates (individual wells) for a single cell line per genotype indicated by black circles. Graph bars indicate mean ± standard deviation.

    Techniques Used: In Vitro, Immunocytochemistry, Derivative Assay, Plasmid Purification, Standard Deviation

    Plp1- targeting ASOs increase Mbp expression and rescue oligodendrocyte numbers in jimpy mice. a , Western blot data showing the level of MBP protein (n=3 mice). b , RT-qPCR data showing the level of Mbp transcript (n=3 mice). c , Western blot data showing the level of MBP (n=3 mice). d , Immunohistochemistry images with hematoxylin counterstain of whole brain sagittal sections showing MBP + myelin. Scale bar, 1mm. e , Quantification of cleaved-caspase 3 + apoptotic cells (n=3 mice). f , Quantification of CC1 + /Olig2 + oligodendrocytes (n=4 mice). g , Quantification of the number of Olig2 + glial lineage cells (n=4 mice). h , Quantification of the number of PDGFRα + /Olig2 + OPCs (n=4 mice). All data collected at 3 weeks post-ASO injection (30μg dose at birth). Individual data points represent the mean value of 4 technical replicates for each biological replicate (individual mice) ( b ) or independent biological replicates (individual mice) ( a, c-h ), indicated by open circles. Graph bars indicate mean ± standard deviation. p-values calculated using one-way ANOVA with Dunnett’s correction for multiple comparisons. p-values stated for p
    Figure Legend Snippet: Plp1- targeting ASOs increase Mbp expression and rescue oligodendrocyte numbers in jimpy mice. a , Western blot data showing the level of MBP protein (n=3 mice). b , RT-qPCR data showing the level of Mbp transcript (n=3 mice). c , Western blot data showing the level of MBP (n=3 mice). d , Immunohistochemistry images with hematoxylin counterstain of whole brain sagittal sections showing MBP + myelin. Scale bar, 1mm. e , Quantification of cleaved-caspase 3 + apoptotic cells (n=3 mice). f , Quantification of CC1 + /Olig2 + oligodendrocytes (n=4 mice). g , Quantification of the number of Olig2 + glial lineage cells (n=4 mice). h , Quantification of the number of PDGFRα + /Olig2 + OPCs (n=4 mice). All data collected at 3 weeks post-ASO injection (30μg dose at birth). Individual data points represent the mean value of 4 technical replicates for each biological replicate (individual mice) ( b ) or independent biological replicates (individual mice) ( a, c-h ), indicated by open circles. Graph bars indicate mean ± standard deviation. p-values calculated using one-way ANOVA with Dunnett’s correction for multiple comparisons. p-values stated for p

    Techniques Used: Expressing, Mouse Assay, Western Blot, Quantitative RT-PCR, Immunohistochemistry, Allele-specific Oligonucleotide, Injection, Standard Deviation

    24) Product Images from "Accumulation of 8,9-unsaturated sterols drives oligodendrocyte formation and remyelination"

    Article Title: Accumulation of 8,9-unsaturated sterols drives oligodendrocyte formation and remyelination

    Journal: Nature

    doi: 10.1038/s41586-018-0360-3

    CYP51 is the functional target by which imidazole antifungals enhance oligodendrocyte formation. a) Azole molecules with varying degrees of potency for mammalian CYP51 inhibition. Throughout, green labels indicate molecules considered active, while red labels indicate inactive molecules. b) Percentage of MBP + oligodendrocytes generated from a second, independent derivation of OPCs (OPC-1) at 72 h following treatment with the indicated concentrations of azoles. n = 4 wells per condition except DMSO (n = 24), with > 1,000 cells analyzed per well. c) GC/MS-based quantitation of lanosterol levels in a second derivation of OPCs (OPC-1) treated 24 h with the indicated azoles at 2.5 μM. n = 2 wells per condition. d, e) GC/MS-based quantitation of cholesterol levels in OPCs (OPC-5 and OPC-1) treated 24 h with the indicated azoles at 2.5 μM. n = 2 wells per condition . f, g) GC/MS-based quantitation of lanosterol levels in OPCs (OPC-5, OPC-1) treated 24 h with the indicated doses of ketoconazole. n = 2 wells per condition. Concentrations shown in panels f and g mirror those shown in Fig. 1c and panel b . h) Percentage of MBP + oligodendrocytes generated from mouse primary OPCs at 72 h following treatment with the indicated imidazole antifungals at 3 μM. n = 4 wells per condition, with > 1,000 cells analyzed per well. i) GC/MS-based quantitation of lanosterol levels in mouse primary OPCs treated 24 h with the indicated imidazole antifungals at 3 μM. n = 2 wells per condition. j) Assessment of oligodendrocyte formation using an alternative image quantitation metric, fold increase in total neurite length. Panel is a re-analysis of data shown in Fig. 1c . n = 4 wells per condition except DMSO (n = 24), with > 1,000 cells analyzed per well. k) Percentage of oligodendrocytes generated from OPCs at 72 h following treatment with ketoconazole (2.5 μM) as measured by PLP1 immunostaining. Left, OPC-5; right, OPC-1. n = 8 wells per condition, with > 1,000 cells analyzed per well. l) LC/MS-based quantitation of lanosterol levels in OPC-5 treated 24 h with ketoconazole at 2.5 μM. n = 2 wells per condition. m) CYP51 mRNA levels measured by RT-qPCR following 96-h treatment with non-targeting or CYP51-targeting pools of cell-permeable siRNAs. n = 2 wells per condition. n ) GC/MS-based quantitation of lanosterol levels in OPC-1 treated 96 h with the indicated pooled siRNA reagents. n = 2 wells per condition. o) Percentage of MBP + oligodendrocytes generated from a second, independent batch of OPCs (OPC-1) at 72 h following treatment with the indicated reagents. n = 3 wells per condition, with > 1,000 cells analyzed per well. p) Percentage of MBP + oligodendrocytes generated from an independent derivation of OPCs at 72 h following treatment with exogenous lanosterol. n = 4 wells per condition except DMSO and Ketoconazole (n = 8), with > 1,000 cells analyzed per well. q) Representative images of OPC-5 cells treated 72 h with the indicated siRNA reagents and lanosterol. Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. All bar graphs indicate mean ± standard deviation, panels b , d, h , i , k , l , o and p are representative of two independent experiments, and all findings have been confirmed in a second independent derivation of OPCs ( Fig. 1 ).
    Figure Legend Snippet: CYP51 is the functional target by which imidazole antifungals enhance oligodendrocyte formation. a) Azole molecules with varying degrees of potency for mammalian CYP51 inhibition. Throughout, green labels indicate molecules considered active, while red labels indicate inactive molecules. b) Percentage of MBP + oligodendrocytes generated from a second, independent derivation of OPCs (OPC-1) at 72 h following treatment with the indicated concentrations of azoles. n = 4 wells per condition except DMSO (n = 24), with > 1,000 cells analyzed per well. c) GC/MS-based quantitation of lanosterol levels in a second derivation of OPCs (OPC-1) treated 24 h with the indicated azoles at 2.5 μM. n = 2 wells per condition. d, e) GC/MS-based quantitation of cholesterol levels in OPCs (OPC-5 and OPC-1) treated 24 h with the indicated azoles at 2.5 μM. n = 2 wells per condition . f, g) GC/MS-based quantitation of lanosterol levels in OPCs (OPC-5, OPC-1) treated 24 h with the indicated doses of ketoconazole. n = 2 wells per condition. Concentrations shown in panels f and g mirror those shown in Fig. 1c and panel b . h) Percentage of MBP + oligodendrocytes generated from mouse primary OPCs at 72 h following treatment with the indicated imidazole antifungals at 3 μM. n = 4 wells per condition, with > 1,000 cells analyzed per well. i) GC/MS-based quantitation of lanosterol levels in mouse primary OPCs treated 24 h with the indicated imidazole antifungals at 3 μM. n = 2 wells per condition. j) Assessment of oligodendrocyte formation using an alternative image quantitation metric, fold increase in total neurite length. Panel is a re-analysis of data shown in Fig. 1c . n = 4 wells per condition except DMSO (n = 24), with > 1,000 cells analyzed per well. k) Percentage of oligodendrocytes generated from OPCs at 72 h following treatment with ketoconazole (2.5 μM) as measured by PLP1 immunostaining. Left, OPC-5; right, OPC-1. n = 8 wells per condition, with > 1,000 cells analyzed per well. l) LC/MS-based quantitation of lanosterol levels in OPC-5 treated 24 h with ketoconazole at 2.5 μM. n = 2 wells per condition. m) CYP51 mRNA levels measured by RT-qPCR following 96-h treatment with non-targeting or CYP51-targeting pools of cell-permeable siRNAs. n = 2 wells per condition. n ) GC/MS-based quantitation of lanosterol levels in OPC-1 treated 96 h with the indicated pooled siRNA reagents. n = 2 wells per condition. o) Percentage of MBP + oligodendrocytes generated from a second, independent batch of OPCs (OPC-1) at 72 h following treatment with the indicated reagents. n = 3 wells per condition, with > 1,000 cells analyzed per well. p) Percentage of MBP + oligodendrocytes generated from an independent derivation of OPCs at 72 h following treatment with exogenous lanosterol. n = 4 wells per condition except DMSO and Ketoconazole (n = 8), with > 1,000 cells analyzed per well. q) Representative images of OPC-5 cells treated 72 h with the indicated siRNA reagents and lanosterol. Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. All bar graphs indicate mean ± standard deviation, panels b , d, h , i , k , l , o and p are representative of two independent experiments, and all findings have been confirmed in a second independent derivation of OPCs ( Fig. 1 ).

    Techniques Used: Functional Assay, Inhibition, Generated, Gas Chromatography-Mass Spectrometry, Quantitation Assay, Immunostaining, Liquid Chromatography with Mass Spectroscopy, Quantitative RT-PCR, Labeling, Standard Deviation

    Effect of independent chemical-genetic and genetic modulators of CYP51, sterol 14 reductase and EBP on oligodendrocyte formation and cholesterol biosynthesis. a, d, g) Percentage of MBP + oligodendrocytes generated from two independent derivation of OPCs at 72 h following treatment with the indicated concentrations of medroxyprogesterone acetate ( a ), 2-methyl ketoconazole ( d ) or TASIN-449 ( g ). n = 4 wells per condition, except DMSO, n = 12 in a and d . In g , for OPC-5, n = 4 except DMSO, n = 7; for OPC-1, n = 3 except DMSO, n = 6. b, e, h) GC/MS-based quantitation of sterol levels in two independent derivations of OPCs treated 24 h with medroxyprogesterone acetate at 10 μM ( a ), 2-methyl ketoconazole at 2.5 μM ( e ) and TASIN-449 at the indicated concentrations ( h ). n = 2 wells per condition. c, f) Rat CYP51 enzymatic activity following treatment with varying concentrations of medroxyprogesterone acetate ( c ) and 2-methyl ketoconazole ( f ) as measured by LC/MS-based quantitation of the CYP51 product FF-MAS. n = 2 independent enzymatic assays. i) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) infected with lentivirus expressing Cas9 and an independent guide RNA targeting EBP (cf. Fig. 2c ). 8 wells per condition, with > 1,000 cells analyzed per well. Two-tailed Student’s t -test, * P = 0.0009. j) Functional validation of CRISPR-based targeting of EBP with a second sgRNA using GC/MS-based quantitation of zymostenol levels. n = 2 wells per condition. k) EBP mRNA levels measured by RT-qPCR in OPCs (OPC-5) infected with lentivirus expressing Cas9 and either of two guide RNAs targeting EBP. One well per condition, with results validated in an independent experiment. l ) Representative images of the oligodendrocyte formation assay shown in Main Figure 2c . Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. All bar graphs indicate mean +/− standard deviation, and panels a, d, g , i and k are representative of two independent experiments.
    Figure Legend Snippet: Effect of independent chemical-genetic and genetic modulators of CYP51, sterol 14 reductase and EBP on oligodendrocyte formation and cholesterol biosynthesis. a, d, g) Percentage of MBP + oligodendrocytes generated from two independent derivation of OPCs at 72 h following treatment with the indicated concentrations of medroxyprogesterone acetate ( a ), 2-methyl ketoconazole ( d ) or TASIN-449 ( g ). n = 4 wells per condition, except DMSO, n = 12 in a and d . In g , for OPC-5, n = 4 except DMSO, n = 7; for OPC-1, n = 3 except DMSO, n = 6. b, e, h) GC/MS-based quantitation of sterol levels in two independent derivations of OPCs treated 24 h with medroxyprogesterone acetate at 10 μM ( a ), 2-methyl ketoconazole at 2.5 μM ( e ) and TASIN-449 at the indicated concentrations ( h ). n = 2 wells per condition. c, f) Rat CYP51 enzymatic activity following treatment with varying concentrations of medroxyprogesterone acetate ( c ) and 2-methyl ketoconazole ( f ) as measured by LC/MS-based quantitation of the CYP51 product FF-MAS. n = 2 independent enzymatic assays. i) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) infected with lentivirus expressing Cas9 and an independent guide RNA targeting EBP (cf. Fig. 2c ). 8 wells per condition, with > 1,000 cells analyzed per well. Two-tailed Student’s t -test, * P = 0.0009. j) Functional validation of CRISPR-based targeting of EBP with a second sgRNA using GC/MS-based quantitation of zymostenol levels. n = 2 wells per condition. k) EBP mRNA levels measured by RT-qPCR in OPCs (OPC-5) infected with lentivirus expressing Cas9 and either of two guide RNAs targeting EBP. One well per condition, with results validated in an independent experiment. l ) Representative images of the oligodendrocyte formation assay shown in Main Figure 2c . Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. All bar graphs indicate mean +/− standard deviation, and panels a, d, g , i and k are representative of two independent experiments.

    Techniques Used: Generated, Gas Chromatography-Mass Spectrometry, Quantitation Assay, Activity Assay, Liquid Chromatography with Mass Spectroscopy, Infection, Expressing, Two Tailed Test, Functional Assay, CRISPR, Quantitative RT-PCR, Tube Formation Assay, Labeling, Immunostaining, Standard Deviation

    Imidazoles inhibit CYP51 to enhance oligodendrocyte formation. a) Rat CYP51 enzymatic activity following treatment with azoles. n = 2 independent enzymatic assays. b) GC/MS-based quantitation of lanosterol levels in OPCs treated with the indicated azoles at 2.5 μM. n = 2 wells per condition. c , f , g) Percentage of MBP + oligodendrocytes generated from OPCs following treatment with azoles ( c ), cell permeable siRNA reagents ( f ), and lanosterol ( g ). n ≥ 4 wells per condition; for exact well counts in all figures, see Statistics and Reproducibility. In f , *, P = 0.0005, two-tailed Student’s t-test. d) Representative images of OPCs treated with the indicated azoles. Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. e) GC/MS-based quantitation of lanosterol levels in OPCs treated with the indicated reagents. n = 2 wells per condition. h ) Structure of lanosterol. All bar graphs indicate mean ± standard deviation. Experiments in c, d, and g are representative of three independent experiments, while b, e, and f are representative of two independent experiments using OPC-5 cells; for validation in an independent derivation of OPCs, see Extended Data Fig. 2 .
    Figure Legend Snippet: Imidazoles inhibit CYP51 to enhance oligodendrocyte formation. a) Rat CYP51 enzymatic activity following treatment with azoles. n = 2 independent enzymatic assays. b) GC/MS-based quantitation of lanosterol levels in OPCs treated with the indicated azoles at 2.5 μM. n = 2 wells per condition. c , f , g) Percentage of MBP + oligodendrocytes generated from OPCs following treatment with azoles ( c ), cell permeable siRNA reagents ( f ), and lanosterol ( g ). n ≥ 4 wells per condition; for exact well counts in all figures, see Statistics and Reproducibility. In f , *, P = 0.0005, two-tailed Student’s t-test. d) Representative images of OPCs treated with the indicated azoles. Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. e) GC/MS-based quantitation of lanosterol levels in OPCs treated with the indicated reagents. n = 2 wells per condition. h ) Structure of lanosterol. All bar graphs indicate mean ± standard deviation. Experiments in c, d, and g are representative of three independent experiments, while b, e, and f are representative of two independent experiments using OPC-5 cells; for validation in an independent derivation of OPCs, see Extended Data Fig. 2 .

    Techniques Used: Activity Assay, Gas Chromatography-Mass Spectrometry, Quantitation Assay, Generated, Two Tailed Test, Labeling, Immunostaining, Standard Deviation

    Inhibiting CYP51, TM7SF2 and EBP is a unifying mechanism for many small-molecule enhancers of oligodendrocyte formation identified by high-throughput screening. a) Percentage of MBP + oligodendrocytes (relative to DMSO control wells) generated from OPCs (OPC-1 derivation) at 72 h following treatment with a library of 3,000 bioactive small molecules, each at 2 μM. Each dot represents the result for one small molecule in the library. Red, imidazole antifungals; blue, clemastine; green, EPZ005687, the top novel hit molecule which is discussed in Extended Data Fig. 7 below. b, c) Percentage of MBP + oligodendrocytes generated from OPCs (Left: OPC-5; Right: OPC-1) at 72 h following treatment with ketoconazole, nine top molecules identified by bioactives screening (green), and nine randomly chosen library members (red) at a uniform dose of 5 μM. n = 4 wells per condition except DMSO and Ketoconazole, n = 12 wells, with > 1,000 cells analyzed per well. d) GC/MS-based quantitation of zymosterol, zymostenol, and 14-dehydrozymostenol levels in a second batch of OPCs treated 24 h with the indicated screening hits and randomly chosen library members at 2 μM. n = 1; for validation in a second derivation of OPCs, see Fig. 3a . Molecules are clustered by enzyme targeted (top labels). e) Percentage of MBP + oligodendrocytes generated from OPCs at 72 h following treatment with the indicated doses of fulvestrant, one of the top 10 HTS hits. n = 4 wells per condition except DMSO, n = 12), with > 1,000 cells analyzed per well. f) GC/MS-based quantitation of lanosterol levels in OPCs treated 24 h with fulvestrant at 2 μM. n = 2 wells per condition. g, h, i) GC/MS-based quantitation of metabolite levels in OPCs treated 24 h with the indicated previously-reported enhancers of oligodendrocyte formation at the following doses: Benztropine, 2 μM; Clemastine, 1 μM; Tamoxifen, 100 nM; U50488, 5 μM; bexarotene, 1 μM; liothyronine, 3 μM. n = 2 wells per condition. j, k) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5 left, OPC-1 right) at 72 h following treatment with the indicated previously-reported enhancers of oligodendrocyte formation. n = 4 wells per condition, except DMSO n = 20 for OPC-5 and n = 12 for OPC-1, with > 1,000 cells analyzed per well. All doses are μM. l) Representative images of OPCs treated 72 h with the indicated small molecules. All treatments in l are at the highest concentration shown in panel j . Scale bar, 100 μm. m) Structures of muscarinic receptor antagonists used in this study. n, q) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5: top, OPC-1: bottom) at 72 h following treatment with ketoconazole or the indicated muscarinic receptor modulators at 2 μM, the concentration used during screening. n = 4 wells per condition except DMSO and Ketoconazole, n = 8, with > 1,000 cells analyzed per well. o) GC/MS-based quantitation of three metabolite levels in OPC-5 OPCs treated 24 h with U50488 (5 μM) or the indicated muscarinic receptor modulators (2 μM). Left, zymostenol; center, cholesterol; right, desmosterol. n = 2 wells per condition. p) Heatmap indicating inhibition of muscarinic receptor isoforms M1, M3, and M5 by the indicated small molecules (2 μM) assayed using GeneBLAzer NFAT- bla CHO-K1 cells. n = 2 wells per condition. r) GC/MS-based quantitation of three metabolite levels in OPC-1 OPCs treated 24 h with clemastine (1 μM) or the indicated muscarinic receptor modulators at 2 μM. n = 2 wells per condition. Left, zymostenol; center, zymosterol; right, cholesterol. p -Fluorohexahydro-sila-difenidol is abbreviated as Sigma H127. All bar graphs indicate mean +/− standard deviation, and panels b , c , e, i, j, k, n and q are representative of two independent experiments.
    Figure Legend Snippet: Inhibiting CYP51, TM7SF2 and EBP is a unifying mechanism for many small-molecule enhancers of oligodendrocyte formation identified by high-throughput screening. a) Percentage of MBP + oligodendrocytes (relative to DMSO control wells) generated from OPCs (OPC-1 derivation) at 72 h following treatment with a library of 3,000 bioactive small molecules, each at 2 μM. Each dot represents the result for one small molecule in the library. Red, imidazole antifungals; blue, clemastine; green, EPZ005687, the top novel hit molecule which is discussed in Extended Data Fig. 7 below. b, c) Percentage of MBP + oligodendrocytes generated from OPCs (Left: OPC-5; Right: OPC-1) at 72 h following treatment with ketoconazole, nine top molecules identified by bioactives screening (green), and nine randomly chosen library members (red) at a uniform dose of 5 μM. n = 4 wells per condition except DMSO and Ketoconazole, n = 12 wells, with > 1,000 cells analyzed per well. d) GC/MS-based quantitation of zymosterol, zymostenol, and 14-dehydrozymostenol levels in a second batch of OPCs treated 24 h with the indicated screening hits and randomly chosen library members at 2 μM. n = 1; for validation in a second derivation of OPCs, see Fig. 3a . Molecules are clustered by enzyme targeted (top labels). e) Percentage of MBP + oligodendrocytes generated from OPCs at 72 h following treatment with the indicated doses of fulvestrant, one of the top 10 HTS hits. n = 4 wells per condition except DMSO, n = 12), with > 1,000 cells analyzed per well. f) GC/MS-based quantitation of lanosterol levels in OPCs treated 24 h with fulvestrant at 2 μM. n = 2 wells per condition. g, h, i) GC/MS-based quantitation of metabolite levels in OPCs treated 24 h with the indicated previously-reported enhancers of oligodendrocyte formation at the following doses: Benztropine, 2 μM; Clemastine, 1 μM; Tamoxifen, 100 nM; U50488, 5 μM; bexarotene, 1 μM; liothyronine, 3 μM. n = 2 wells per condition. j, k) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5 left, OPC-1 right) at 72 h following treatment with the indicated previously-reported enhancers of oligodendrocyte formation. n = 4 wells per condition, except DMSO n = 20 for OPC-5 and n = 12 for OPC-1, with > 1,000 cells analyzed per well. All doses are μM. l) Representative images of OPCs treated 72 h with the indicated small molecules. All treatments in l are at the highest concentration shown in panel j . Scale bar, 100 μm. m) Structures of muscarinic receptor antagonists used in this study. n, q) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5: top, OPC-1: bottom) at 72 h following treatment with ketoconazole or the indicated muscarinic receptor modulators at 2 μM, the concentration used during screening. n = 4 wells per condition except DMSO and Ketoconazole, n = 8, with > 1,000 cells analyzed per well. o) GC/MS-based quantitation of three metabolite levels in OPC-5 OPCs treated 24 h with U50488 (5 μM) or the indicated muscarinic receptor modulators (2 μM). Left, zymostenol; center, cholesterol; right, desmosterol. n = 2 wells per condition. p) Heatmap indicating inhibition of muscarinic receptor isoforms M1, M3, and M5 by the indicated small molecules (2 μM) assayed using GeneBLAzer NFAT- bla CHO-K1 cells. n = 2 wells per condition. r) GC/MS-based quantitation of three metabolite levels in OPC-1 OPCs treated 24 h with clemastine (1 μM) or the indicated muscarinic receptor modulators at 2 μM. n = 2 wells per condition. Left, zymostenol; center, zymosterol; right, cholesterol. p -Fluorohexahydro-sila-difenidol is abbreviated as Sigma H127. All bar graphs indicate mean +/− standard deviation, and panels b , c , e, i, j, k, n and q are representative of two independent experiments.

    Techniques Used: High Throughput Screening Assay, Generated, Gas Chromatography-Mass Spectrometry, Quantitation Assay, Concentration Assay, Inhibition, Standard Deviation

    Effect of 8,9-unsaturated sterols on oligodendrocyte formation. a) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) at 72 h following treatment with methyl β-cyclodextrin (1 mM) for 30 min at 37 °C. n = 8 wells per condition, with > 1,000 cells analyzed per well. b) GC/MS-based quantitation of cholesterol (left) and desmosterol (right) levels in OPCs (OPC-5) treated with methyl β-cyclodextrin (Me-β-CD) at 1 mM or ketoconazole at 2.5 μM. n = 2 wells per condition. c, d) Percentage of MBP + oligodendrocytes generated from OPC-1 ( c ) and OPC-5 cells ( d ) at 72 h following treatment with the indicated purified sterol intermediates. n = 4 wells per condition, except n = 8 for DMSO and ketoconazole, with > 1,000 cells analyzed per well. Green text highlights metabolites that accumulate after treatments that enhance oligodendrocyte formation ( Fig. 2e , Extended Data Fig. 3c ). e) Percentage of MBP+ oligodendrocytes generated from OPC­1 at 72 h following treatment with MAS-412 and MAS-414. n = 4 wells per condition, with > 1,000 cells analyzed per well. f) Representative images of OPC­5 cells treated 72 h with DMSO, MAS-412, or MAS-414 (3 μM). Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. g) Percentage of MBP + oligodendrocytes generated from OPC-1 at 72 h following treatment with 2,2-dimethyl-zymosterol. n = 4 wells per condition except DMSO (n = 12), with > 1,000 cells analyzed per well. h) Representative images of OPC-5 cells treated 72 h with vehicle and 2,2-dimethyl-zymosterol (2.5 μM). Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. i) Percentage of MBP+ oligodendrocytes generated from OPC-5 (left) and OPC-1 (right) at 72 h following treatment with FF-MAS or T-MAS. n = 4 wells per condition except DMSO and Ketoconazole (n = 8), with > 1,000 cells analyzed per well. j) Percentage of MBP + oligodendrocytes generated from OPC-5 and OPC-1 OPCs at 72 h following treatment with the indicated concentrations of cholesterol. n = 8 wells per condition, with > 1,000 cells analyzed per well. k, l) Percentage of MBP + oligodendrocytes generated from OPC-5 and OPC-1 at 72 h following treatment with the indicated concentrations of sterols that are structurally identical aside from the presence or absence of the 8,9 double bond (structures in panel o ). n ≥ 3 wells per condition (see dot plots as replicate values vary by condition), with > 1,000 cells analyzed per well. m) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) at 72 h following treatment with the indicated small molecules or combinations of small molecules (ketoconazole, 2.5 μM; Ro 48–8071, 11 nM; liothyronine, 3 μM). n = 3 wells per condition, except DMSO n = 11, ketoconazole n = 13, liothyronine n = 8 liothyronine + Ro 48–8071 n = 4, with > 1,000 cells analyzed per well. n) GC/MS-based quantitation of lanosterol levels in OPCs (OPC-5) treated 24 h with the indicated small molecules or combinations of small molecules at concentrations stated in m. n = 2 wells per condition. o) Structures of zymostenol, 8,9-dehydrocholesterol, 5α-cholestanol, and cholesterol. p) Total cell number as measured by counting of DAPI+ nuclei in the experiment presented in panel m . q, r) Percentage of MBP+ oligodendrocytes generated from OPCs (OPC­5 and OPC-1) at 72 h following treatment with the indicated small molecules or combinations of small molecules in two independent batches of OPCs (ketoconazole, 2.5 μM; MAS­412, 5 μM). In q , n = 16 for DMSO, 8 for Ketoconazole, and 4 for remaining bars. In r , n = 8 wells per condition. s) Luciferase reporter assays were used to assess if 2,2-Dimethylzymosterol (5 μM), Ketoconazole (2.5 μM), and TASIN-1 (250 nM) modulate human ERα, GR, LXRβ, NFkB, NRF2, PGR, PPARδ, PPARγ, RARα, RARγ, RXRα, RXRβ, TRα, TRβ and VDR transcriptional activity in agonist mode and ERRα, RORα and RORγ in inverse-agonist mode. n = 2 wells per condition and n = 3 wells per positive control conditions. t) Effects of sterols (2,2-dimethylzymosterol 5 μM, FF-MAS 10 μM) and small molecules (Ketoconazole 2.5 μM, TASIN-1 100 nM) on the NR2F1-mediated activation of a NGFI-A promoter driven luciferase reporter. n = 2 wells per condition. u) Effects of 2,2-dimethylzymosterol (5 μM) on NR2C2-mediated activation of a NGFI-A promoter driven luciferase reporter in comparison to cells transfected with reporter only, untreated, or treated with a previously reported positive control (all-trans retinoic acid, ATRA, 5 μM). n = 2 wells per condition. v) LSS, DHCR7, LDLR mRNA levels measured by RT-qPCR following 24 h treatment with DMSO, Mevastatin (2.5 μM), Ro 48–8071 (500 nM), Ketoconazole (2.5 μM), TASIN-1 (100 nM), or Amorolfine (100 nM). n = 2 wells. All bar graphs indicate mean +/− standard deviation, and panels a-n , and t-v are representative of two independent experiments.
    Figure Legend Snippet: Effect of 8,9-unsaturated sterols on oligodendrocyte formation. a) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) at 72 h following treatment with methyl β-cyclodextrin (1 mM) for 30 min at 37 °C. n = 8 wells per condition, with > 1,000 cells analyzed per well. b) GC/MS-based quantitation of cholesterol (left) and desmosterol (right) levels in OPCs (OPC-5) treated with methyl β-cyclodextrin (Me-β-CD) at 1 mM or ketoconazole at 2.5 μM. n = 2 wells per condition. c, d) Percentage of MBP + oligodendrocytes generated from OPC-1 ( c ) and OPC-5 cells ( d ) at 72 h following treatment with the indicated purified sterol intermediates. n = 4 wells per condition, except n = 8 for DMSO and ketoconazole, with > 1,000 cells analyzed per well. Green text highlights metabolites that accumulate after treatments that enhance oligodendrocyte formation ( Fig. 2e , Extended Data Fig. 3c ). e) Percentage of MBP+ oligodendrocytes generated from OPC­1 at 72 h following treatment with MAS-412 and MAS-414. n = 4 wells per condition, with > 1,000 cells analyzed per well. f) Representative images of OPC­5 cells treated 72 h with DMSO, MAS-412, or MAS-414 (3 μM). Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. g) Percentage of MBP + oligodendrocytes generated from OPC-1 at 72 h following treatment with 2,2-dimethyl-zymosterol. n = 4 wells per condition except DMSO (n = 12), with > 1,000 cells analyzed per well. h) Representative images of OPC-5 cells treated 72 h with vehicle and 2,2-dimethyl-zymosterol (2.5 μM). Nuclei are labeled with DAPI (blue), and oligodendrocytes are indicated by immunostaining for myelin basic protein (green). Scale bar, 100 μm. i) Percentage of MBP+ oligodendrocytes generated from OPC-5 (left) and OPC-1 (right) at 72 h following treatment with FF-MAS or T-MAS. n = 4 wells per condition except DMSO and Ketoconazole (n = 8), with > 1,000 cells analyzed per well. j) Percentage of MBP + oligodendrocytes generated from OPC-5 and OPC-1 OPCs at 72 h following treatment with the indicated concentrations of cholesterol. n = 8 wells per condition, with > 1,000 cells analyzed per well. k, l) Percentage of MBP + oligodendrocytes generated from OPC-5 and OPC-1 at 72 h following treatment with the indicated concentrations of sterols that are structurally identical aside from the presence or absence of the 8,9 double bond (structures in panel o ). n ≥ 3 wells per condition (see dot plots as replicate values vary by condition), with > 1,000 cells analyzed per well. m) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-5) at 72 h following treatment with the indicated small molecules or combinations of small molecules (ketoconazole, 2.5 μM; Ro 48–8071, 11 nM; liothyronine, 3 μM). n = 3 wells per condition, except DMSO n = 11, ketoconazole n = 13, liothyronine n = 8 liothyronine + Ro 48–8071 n = 4, with > 1,000 cells analyzed per well. n) GC/MS-based quantitation of lanosterol levels in OPCs (OPC-5) treated 24 h with the indicated small molecules or combinations of small molecules at concentrations stated in m. n = 2 wells per condition. o) Structures of zymostenol, 8,9-dehydrocholesterol, 5α-cholestanol, and cholesterol. p) Total cell number as measured by counting of DAPI+ nuclei in the experiment presented in panel m . q, r) Percentage of MBP+ oligodendrocytes generated from OPCs (OPC­5 and OPC-1) at 72 h following treatment with the indicated small molecules or combinations of small molecules in two independent batches of OPCs (ketoconazole, 2.5 μM; MAS­412, 5 μM). In q , n = 16 for DMSO, 8 for Ketoconazole, and 4 for remaining bars. In r , n = 8 wells per condition. s) Luciferase reporter assays were used to assess if 2,2-Dimethylzymosterol (5 μM), Ketoconazole (2.5 μM), and TASIN-1 (250 nM) modulate human ERα, GR, LXRβ, NFkB, NRF2, PGR, PPARδ, PPARγ, RARα, RARγ, RXRα, RXRβ, TRα, TRβ and VDR transcriptional activity in agonist mode and ERRα, RORα and RORγ in inverse-agonist mode. n = 2 wells per condition and n = 3 wells per positive control conditions. t) Effects of sterols (2,2-dimethylzymosterol 5 μM, FF-MAS 10 μM) and small molecules (Ketoconazole 2.5 μM, TASIN-1 100 nM) on the NR2F1-mediated activation of a NGFI-A promoter driven luciferase reporter. n = 2 wells per condition. u) Effects of 2,2-dimethylzymosterol (5 μM) on NR2C2-mediated activation of a NGFI-A promoter driven luciferase reporter in comparison to cells transfected with reporter only, untreated, or treated with a previously reported positive control (all-trans retinoic acid, ATRA, 5 μM). n = 2 wells per condition. v) LSS, DHCR7, LDLR mRNA levels measured by RT-qPCR following 24 h treatment with DMSO, Mevastatin (2.5 μM), Ro 48–8071 (500 nM), Ketoconazole (2.5 μM), TASIN-1 (100 nM), or Amorolfine (100 nM). n = 2 wells. All bar graphs indicate mean +/− standard deviation, and panels a-n , and t-v are representative of two independent experiments.

    Techniques Used: Generated, Gas Chromatography-Mass Spectrometry, Quantitation Assay, Purification, Labeling, Immunostaining, Luciferase, Activity Assay, Positive Control, Activation Assay, Transfection, Quantitative RT-PCR, Standard Deviation

    Effect of combinations of small-molecule treatments on oligodendrocyte formation, and ability of oligodendrocytes to track along and wrap electrospun microfibers after single small molecule treatments. a, b) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-1: Left, OPC-5: Right) at 72 h following treatment with the indicated combinations of liothyronine and enhancers of oligodendrocyte formation. Unless noted, the following concentrations were used: ketoconazole, 2.5 μM; benztropine, 2 μM; clemastine 2 μM; tamoxifen 200 nM; liothyronine, 3 μM. n = 4 wells per treatment condition, with > 1,000 cells analyzed per well. Lio = liothyronine. c, d) Percentage of MBP + oligodendrocytes generated from OPCs at 72 h following treatment with the indicated combinations of ketoconazole and enhancers of oligodendrocyte formation. n = 4 wells per treatment condition, with > 1,000 cells analyzed per well. Keto = ketoconazole. e) Representative images of OPCs treated 72 h with the indicated small molecules. Small-molecule concentrations are as in panel a . Scale bar, 100 μm. f) Fold-increase of MBP+ oligodendrocytes following plating of OPCs (OPC-5) onto microfibers and treatment for 14 days with the indicated pathway modulators. n = 2 wells per condition, except DMSO, n = 4. g) In an independent experiment, OPCs (OPC-5) were plated onto microfibers, treated with small molecules for 4 days, and fixed and stained after 14 days. The extent to which MBP+ oligodendrocytes tracked along the microfiber substrate was measured. n = 2 wells per condition. h) Total DAPI+ cell number in the experiment described in g . i) Representative images highlighting tracking along the microfiber substrate. Each image is a montage of 4 separate images within the same well. Green, MBP. Scale bar, 100 μm. j) High-resolution images of MBP+ oligodendrocytes tracking along microfibers. Green, MBP; Blue, DAPI. Ketoconazole, 2.5 μM. Scale bar, 50 μm. k) Confocal imaging of OPCs seeded onto aligned microfibers and treated 14 days with ketoconazole (2.5 μM). The plane of the cross-section is highlighted in yellow and the cross-section, in which green fluorescence appears to encircle several microfibers, is shown in the bottom panel. Green, MBP; Blue, DAPI. All bar graphs indicate mean +/− standard deviation, and panels a-d are representative of two independent experiments.
    Figure Legend Snippet: Effect of combinations of small-molecule treatments on oligodendrocyte formation, and ability of oligodendrocytes to track along and wrap electrospun microfibers after single small molecule treatments. a, b) Percentage of MBP + oligodendrocytes generated from OPCs (OPC-1: Left, OPC-5: Right) at 72 h following treatment with the indicated combinations of liothyronine and enhancers of oligodendrocyte formation. Unless noted, the following concentrations were used: ketoconazole, 2.5 μM; benztropine, 2 μM; clemastine 2 μM; tamoxifen 200 nM; liothyronine, 3 μM. n = 4 wells per treatment condition, with > 1,000 cells analyzed per well. Lio = liothyronine. c, d) Percentage of MBP + oligodendrocytes generated from OPCs at 72 h following treatment with the indicated combinations of ketoconazole and enhancers of oligodendrocyte formation. n = 4 wells per treatment condition, with > 1,000 cells analyzed per well. Keto = ketoconazole. e) Representative images of OPCs treated 72 h with the indicated small molecules. Small-molecule concentrations are as in panel a . Scale bar, 100 μm. f) Fold-increase of MBP+ oligodendrocytes following plating of OPCs (OPC-5) onto microfibers and treatment for 14 days with the indicated pathway modulators. n = 2 wells per condition, except DMSO, n = 4. g) In an independent experiment, OPCs (OPC-5) were plated onto microfibers, treated with small molecules for 4 days, and fixed and stained after 14 days. The extent to which MBP+ oligodendrocytes tracked along the microfiber substrate was measured. n = 2 wells per condition. h) Total DAPI+ cell number in the experiment described in g . i) Representative images highlighting tracking along the microfiber substrate. Each image is a montage of 4 separate images within the same well. Green, MBP. Scale bar, 100 μm. j) High-resolution images of MBP+ oligodendrocytes tracking along microfibers. Green, MBP; Blue, DAPI. Ketoconazole, 2.5 μM. Scale bar, 50 μm. k) Confocal imaging of OPCs seeded onto aligned microfibers and treated 14 days with ketoconazole (2.5 μM). The plane of the cross-section is highlighted in yellow and the cross-section, in which green fluorescence appears to encircle several microfibers, is shown in the bottom panel. Green, MBP; Blue, DAPI. All bar graphs indicate mean +/− standard deviation, and panels a-d are representative of two independent experiments.

    Techniques Used: Generated, Staining, Imaging, Fluorescence, Standard Deviation

    25) Product Images from "FGF1 improves functional recovery through inducing PRDX1 to regulate autophagy and anti‐ROS after spinal cord injury, et al. FGF1 improves functional recovery through inducing PRDX1 to reduce autophagy and ROS after spinal cord injury"

    Article Title: FGF1 improves functional recovery through inducing PRDX1 to regulate autophagy and anti‐ROS after spinal cord injury, et al. FGF1 improves functional recovery through inducing PRDX1 to reduce autophagy and ROS after spinal cord injury

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13566

    AAV ‐ FGF 1 promoted neuroprotection, axon regeneration and remyelination after spinal cord injury. A, Representative Western blot result of MBP , ace‐tubulin, GAP 43 and FGF 1 at 14 dpi in each group. B‐E, Quantification of Western blot data from A, *** P
    Figure Legend Snippet: AAV ‐ FGF 1 promoted neuroprotection, axon regeneration and remyelination after spinal cord injury. A, Representative Western blot result of MBP , ace‐tubulin, GAP 43 and FGF 1 at 14 dpi in each group. B‐E, Quantification of Western blot data from A, *** P

    Techniques Used: Western Blot

    26) Product Images from "Luoyutong Treatment Promotes Functional Recovery and Neuronal Plasticity after Cerebral Ischemia-Reperfusion Injury in Rats"

    Article Title: Luoyutong Treatment Promotes Functional Recovery and Neuronal Plasticity after Cerebral Ischemia-Reperfusion Injury in Rats

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    doi: 10.1155/2015/369021

    Schematic representation of the experimental procedures. (Luoyutong: LYT; middle cerebral artery occlusion: MCAO; 2,3,5-triphenyltetrazolium chloride: TTC; microtubule associated protein: MAP-2; myelin basic protein: MBP; brain derived neurotrophic factor: BDNF; basic fibroblast growth factor: b-FGF; neuron-specific nuclear protein: NeuN; neurofilament 200: NF200).
    Figure Legend Snippet: Schematic representation of the experimental procedures. (Luoyutong: LYT; middle cerebral artery occlusion: MCAO; 2,3,5-triphenyltetrazolium chloride: TTC; microtubule associated protein: MAP-2; myelin basic protein: MBP; brain derived neurotrophic factor: BDNF; basic fibroblast growth factor: b-FGF; neuron-specific nuclear protein: NeuN; neurofilament 200: NF200).

    Techniques Used: Derivative Assay

    LYT (Luoyutong) induced upregulation of MBP (myelin basic protein) expression after MCAO (middle cerebral artery occlusion). Western blot detection and quantitative analysis of (a) MBP expression 3 days after MCAO and (c) MBP expression 14 days after MCAO. (b) Representative immunofluorescence images showing colocalization of MBP (red) and NF200 (neurofilament 200) (green) in the cortex. Blue DAPI staining indicates the nuclei. n = 3. ∗∗ P
    Figure Legend Snippet: LYT (Luoyutong) induced upregulation of MBP (myelin basic protein) expression after MCAO (middle cerebral artery occlusion). Western blot detection and quantitative analysis of (a) MBP expression 3 days after MCAO and (c) MBP expression 14 days after MCAO. (b) Representative immunofluorescence images showing colocalization of MBP (red) and NF200 (neurofilament 200) (green) in the cortex. Blue DAPI staining indicates the nuclei. n = 3. ∗∗ P

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Staining

    27) Product Images from "TLR4 response mediates ethanol-induced neurodevelopment alterations in a model of fetal alcohol spectrum disorders"

    Article Title: TLR4 response mediates ethanol-induced neurodevelopment alterations in a model of fetal alcohol spectrum disorders

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-017-0918-2

    Role of TLR4 in the expression of several proteins induced by prenatal and postnatal ethanol exposure. Data represent the immunoblot analysis of PLP, MBP, synaptotagmin, synapsin IIa, Tuj-1, CD11b, MHC-II, caspase 3 (active fragment of 17 kDa), and the ELISA analysis of MAP-2 from the cerebral cortices of the WT and TLR4-KO pups on PND 0, 20, and 66 exposed (E), or not (C), to ethanol during the embryonic and postnatal periods. A representative immunoblot of each protein is shown. PPEE prenatal and postnatal ethanol exposure. Data represent mean ± SEM, n = 8 mice/group. * p
    Figure Legend Snippet: Role of TLR4 in the expression of several proteins induced by prenatal and postnatal ethanol exposure. Data represent the immunoblot analysis of PLP, MBP, synaptotagmin, synapsin IIa, Tuj-1, CD11b, MHC-II, caspase 3 (active fragment of 17 kDa), and the ELISA analysis of MAP-2 from the cerebral cortices of the WT and TLR4-KO pups on PND 0, 20, and 66 exposed (E), or not (C), to ethanol during the embryonic and postnatal periods. A representative immunoblot of each protein is shown. PPEE prenatal and postnatal ethanol exposure. Data represent mean ± SEM, n = 8 mice/group. * p

    Techniques Used: Expressing, Plasmid Purification, Enzyme-linked Immunosorbent Assay, Mouse Assay

    Role of TLR4 in prenatal and postnatal ethanol exposure in PLP, MBP, and Iba-1 immunoreactivity in the cortices of the WT and TLR4-KO pups. Arrows show examples of microglial activation. The scale bar is 50 μm. Bars represent the quantification values of PLP, MBP, and Iba-1 immunoreactivity, expressed as the thresholded area occupied by specific staining in the cortices of the WT and TLR4-KO mice treated, or not, prenatally and postnatally with ethanol (PPEE). Results are given as means ± SEM ( n = 5). p
    Figure Legend Snippet: Role of TLR4 in prenatal and postnatal ethanol exposure in PLP, MBP, and Iba-1 immunoreactivity in the cortices of the WT and TLR4-KO pups. Arrows show examples of microglial activation. The scale bar is 50 μm. Bars represent the quantification values of PLP, MBP, and Iba-1 immunoreactivity, expressed as the thresholded area occupied by specific staining in the cortices of the WT and TLR4-KO mice treated, or not, prenatally and postnatally with ethanol (PPEE). Results are given as means ± SEM ( n = 5). p

    Techniques Used: Plasmid Purification, Activation Assay, Staining, Mouse Assay

    28) Product Images from "White matter alterations in Alzheimer’s disease without concomitant pathologies"

    Article Title: White matter alterations in Alzheimer’s disease without concomitant pathologies

    Journal: Neuropathology and Applied Neurobiology

    doi: 10.1111/nan.12618

    Immunohistochemistry to myelin markers PLP1, CNPase and MBP in the centrum semi‐ovale of the frontal lobe in MA ( A , E , I ), and in cases with AD without co‐morbidities at stages ADI–II/0‐A (ADI–II) ( B , F , J ), ADIII–IV/0‐C (ADIII–IV) ( C , G , K ) and ADV–VI/B–C (ADV–VI) ( D , H , L ). Representative images show reduced immunoreactivity with disease progression, and small PLP1‐ and CNPase‐immunoreactive dots in ADV–VI. Paraffin sections, slightly counterstained with haematoxylin; bar = 50 µm.
    Figure Legend Snippet: Immunohistochemistry to myelin markers PLP1, CNPase and MBP in the centrum semi‐ovale of the frontal lobe in MA ( A , E , I ), and in cases with AD without co‐morbidities at stages ADI–II/0‐A (ADI–II) ( B , F , J ), ADIII–IV/0‐C (ADIII–IV) ( C , G , K ) and ADV–VI/B–C (ADV–VI) ( D , H , L ). Representative images show reduced immunoreactivity with disease progression, and small PLP1‐ and CNPase‐immunoreactive dots in ADV–VI. Paraffin sections, slightly counterstained with haematoxylin; bar = 50 µm.

    Techniques Used: Immunohistochemistry

    29) Product Images from "TREM2 deficiency eliminates TREM2+ inflammatory macrophages and ameliorates pathology in Alzheimer’s disease mouse models"

    Article Title: TREM2 deficiency eliminates TREM2+ inflammatory macrophages and ameliorates pathology in Alzheimer’s disease mouse models

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20142322

    TREM2 is expressed in plaque-associated myeloid cells. (a) In situ hybridization with TREM2 probes colocalized with Iba1 ( n = 2). (b) X-gal staining of brain tissue from 4-mo-old APPPS1; Trem2 LacZ/+ mice colocalized with fluorescent IHC for Iba1 and 6E10 ( n = 3). (c–f) Confocal microscopy was used to assess TREM2 colocalization with 6E10 + plaque-associated myeloid cells (c; Iba1), astrocytes (d; GFAP), neurons (e; MAP2), or oligodendrocytes (f; MBP; n = 8). At least two independent experiments were performed for all analyses. Bars: (a) 20 µm; (b–f) 50 µm.
    Figure Legend Snippet: TREM2 is expressed in plaque-associated myeloid cells. (a) In situ hybridization with TREM2 probes colocalized with Iba1 ( n = 2). (b) X-gal staining of brain tissue from 4-mo-old APPPS1; Trem2 LacZ/+ mice colocalized with fluorescent IHC for Iba1 and 6E10 ( n = 3). (c–f) Confocal microscopy was used to assess TREM2 colocalization with 6E10 + plaque-associated myeloid cells (c; Iba1), astrocytes (d; GFAP), neurons (e; MAP2), or oligodendrocytes (f; MBP; n = 8). At least two independent experiments were performed for all analyses. Bars: (a) 20 µm; (b–f) 50 µm.

    Techniques Used: In Situ Hybridization, Staining, Mouse Assay, Immunohistochemistry, Confocal Microscopy

    30) Product Images from "Transplantation of human induced cerebellar granular-like cells improves motor functions in a novel mouse model of cerebellar ataxia"

    Article Title: Transplantation of human induced cerebellar granular-like cells improves motor functions in a novel mouse model of cerebellar ataxia

    Journal: American Journal of Translational Research

    doi:

    hiCGCs expressed human neural subtype and cerebellum specific markers (En1, Ath1, Zic1), neuronal subtype markers GABA and vGluT1. hiCGCs were negative for MBP and GFAP. Nuclei were stained with 4, 6-diamidino-2-phenylindole (DAPI).
    Figure Legend Snippet: hiCGCs expressed human neural subtype and cerebellum specific markers (En1, Ath1, Zic1), neuronal subtype markers GABA and vGluT1. hiCGCs were negative for MBP and GFAP. Nuclei were stained with 4, 6-diamidino-2-phenylindole (DAPI).

    Techniques Used: Staining

    31) Product Images from "Involvement of Atm and Trp53 in neural cell loss due to Terf2 inactivation during mouse brain development"

    Article Title: Involvement of Atm and Trp53 in neural cell loss due to Terf2 inactivation during mouse brain development

    Journal: Histochemistry and Cell Biology

    doi: 10.1007/s00418-017-1591-3

    Defective glia network was formed in the mutant brain. a Increased immunoreactivity of GFAP in the Terf2 mutant brain. The Terf2 -null brains in both Atm- and Trp53- deficient backgrounds showed the high level of GFAP immuno-staining compared to that of the control brain, which is a common feature found in the brain of genomic instability animal models. The corpus callosum, which is a glia-rich structure, was not defined in the Terf2 -null brain ( (Terf2;Trp53) Nes - Cre , Terf2 Emx1 - Cre ;Atm − / − and (Terf2;Trp53) Emx1 - Cre ). The (Terf2;Lig4;Trp53) Emx1 - Cre brain showed partial recovery of the corpus callosum. The red dotted lines demarcate the normal corpus callosum in the control brain and the partially restored corpus callosum in the (Terf2;Lig4;Trp53) Emx1 - Cre brain. The size of GFAP immuno-positive cells in the Terf2/Trp53 double null brains, not in both Terf2/Atm double and Terf2/Lig4/Trp53 triple deficient brains, was bigger than that of controls ( red arrows ). The scale bar is 200 μm. b Defective oligodendrocyte network in the Terf2 mutant brain. Oligodendrocytes were detected by two different markers: MBP (myelin basic protein) and Myelin-PLP (proteolipid protein). The Terf2 ctrl cerebral cortex displayed a rich network of oligodendrocytes, particularly in the corpus callosum. However, the Terf2 conditional knockout brains in all different genetic backgrounds showed a dramatic reduction of immunopositivity to MBP/Myelin-PLP, and agenesis of the corpus callosum ( red arrows ), in contrast to the increased astrocyte population in the Terf2 -null brains (A panel above ). And the genetic combination with Atm or Trp53 deficiency could not improve this defect. However, Lig4 inactivation resulted in partial recovery of the corpus callosum in the (Terf2;Trp53) Emx1 - Cr e brain. The corpus callosum shown in the lower panel was the posterior part of the corpus callosum. The scale bars are 100 μm
    Figure Legend Snippet: Defective glia network was formed in the mutant brain. a Increased immunoreactivity of GFAP in the Terf2 mutant brain. The Terf2 -null brains in both Atm- and Trp53- deficient backgrounds showed the high level of GFAP immuno-staining compared to that of the control brain, which is a common feature found in the brain of genomic instability animal models. The corpus callosum, which is a glia-rich structure, was not defined in the Terf2 -null brain ( (Terf2;Trp53) Nes - Cre , Terf2 Emx1 - Cre ;Atm − / − and (Terf2;Trp53) Emx1 - Cre ). The (Terf2;Lig4;Trp53) Emx1 - Cre brain showed partial recovery of the corpus callosum. The red dotted lines demarcate the normal corpus callosum in the control brain and the partially restored corpus callosum in the (Terf2;Lig4;Trp53) Emx1 - Cre brain. The size of GFAP immuno-positive cells in the Terf2/Trp53 double null brains, not in both Terf2/Atm double and Terf2/Lig4/Trp53 triple deficient brains, was bigger than that of controls ( red arrows ). The scale bar is 200 μm. b Defective oligodendrocyte network in the Terf2 mutant brain. Oligodendrocytes were detected by two different markers: MBP (myelin basic protein) and Myelin-PLP (proteolipid protein). The Terf2 ctrl cerebral cortex displayed a rich network of oligodendrocytes, particularly in the corpus callosum. However, the Terf2 conditional knockout brains in all different genetic backgrounds showed a dramatic reduction of immunopositivity to MBP/Myelin-PLP, and agenesis of the corpus callosum ( red arrows ), in contrast to the increased astrocyte population in the Terf2 -null brains (A panel above ). And the genetic combination with Atm or Trp53 deficiency could not improve this defect. However, Lig4 inactivation resulted in partial recovery of the corpus callosum in the (Terf2;Trp53) Emx1 - Cr e brain. The corpus callosum shown in the lower panel was the posterior part of the corpus callosum. The scale bars are 100 μm

    Techniques Used: Mutagenesis, Immunostaining, Plasmid Purification, Knock-Out

    32) Product Images from "Gene therapy targeting oligodendrocytes provides therapeutic benefit in a leukodystrophy model"

    Article Title: Gene therapy targeting oligodendrocytes provides therapeutic benefit in a leukodystrophy model

    Journal: Brain

    doi: 10.1093/brain/aww351

    Re-establishment of oligodendrocyte network connectivity following Cx47 gene delivery with the AAV.MBP.Cx47myc vector . ( A ) Oligodendrocytes were identified by their transgenic expression of green fluorescence in all Cx32/Cx47 double knockout mice (see methods). ( B ) Representative membrane current recording from an oligodendrocyte. ( C and D ) Examples of Lucifer yellow (visualized red) injection into an oligodendrocyte (asterisk) in the corpus callosum in untreated double knockout mouse ( C ) and in an AAV.MBP.Cx47myc injected mouse ( D ). Whereas no transfer to another cell is noticed 30 thinsp;min after injection in the double knockout, a red fluorescent signal in at least two adjacent cells (arrows) is observed in the treated mouse, suggesting establishment of network connectivity. Quantification of multiple experiments from positive control mice (TG plus; are double knockout expressing transgenically Cx32 in all oligodendrocytes, see methods) ( n thinsp; equals; thinsp; 4 mice, four slices), from untreated double knockout mice ( n thinsp; equals; thinsp; 5 mice, six slices) and from treated double knockout mice ( n thinsp; equals; thinsp; 6 mice, six slices) shows that the percentage of network coupled oligodendrocytes per total cells ( E ) has markedly increased in the treated mice (85.7 percnt;), compared to 20 percnt; of untreated double knockout mice, and 100 percnt; of positive control TG plus; mice. ( F ) On average there is dye transfer to significantly more oligodendrocytes (number of couple cells) in treated (2.29 thinsp; plusmn; thinsp;0.69) compared to untreated (0.4 thinsp; plusmn; thinsp;0.690.23) mice ( P thinsp; equals; thinsp; 0.048, unpaired Student t -test), reaching the levels of transgenic mouse network connectivity (2.25 thinsp; plusmn; thinsp;0.63). dKO thinsp; equals; thinsp;double knockout.
    Figure Legend Snippet: Re-establishment of oligodendrocyte network connectivity following Cx47 gene delivery with the AAV.MBP.Cx47myc vector . ( A ) Oligodendrocytes were identified by their transgenic expression of green fluorescence in all Cx32/Cx47 double knockout mice (see methods). ( B ) Representative membrane current recording from an oligodendrocyte. ( C and D ) Examples of Lucifer yellow (visualized red) injection into an oligodendrocyte (asterisk) in the corpus callosum in untreated double knockout mouse ( C ) and in an AAV.MBP.Cx47myc injected mouse ( D ). Whereas no transfer to another cell is noticed 30 thinsp;min after injection in the double knockout, a red fluorescent signal in at least two adjacent cells (arrows) is observed in the treated mouse, suggesting establishment of network connectivity. Quantification of multiple experiments from positive control mice (TG plus; are double knockout expressing transgenically Cx32 in all oligodendrocytes, see methods) ( n thinsp; equals; thinsp; 4 mice, four slices), from untreated double knockout mice ( n thinsp; equals; thinsp; 5 mice, six slices) and from treated double knockout mice ( n thinsp; equals; thinsp; 6 mice, six slices) shows that the percentage of network coupled oligodendrocytes per total cells ( E ) has markedly increased in the treated mice (85.7 percnt;), compared to 20 percnt; of untreated double knockout mice, and 100 percnt; of positive control TG plus; mice. ( F ) On average there is dye transfer to significantly more oligodendrocytes (number of couple cells) in treated (2.29 thinsp; plusmn; thinsp;0.69) compared to untreated (0.4 thinsp; plusmn; thinsp;0.690.23) mice ( P thinsp; equals; thinsp; 0.048, unpaired Student t -test), reaching the levels of transgenic mouse network connectivity (2.25 thinsp; plusmn; thinsp;0.63). dKO thinsp; equals; thinsp;double knockout.

    Techniques Used: Plasmid Preparation, Transgenic Assay, Expressing, Fluorescence, Double Knockout, Mouse Assay, Injection, Positive Control

    AAV.MBP.Cx47myc leads to expression of Cx47 and gap junction formation in oligodendrocytes of Cx47 knockout mice. Double immunostaining for Cx47 (red) and either the myc tag ( A ndash;C ) or the oligodendrocyte marker CC1 ( D ndash;F ) (green) shows Cx47 expression and formation of gap junction (GJ) like plaques (open arrowheads) at the pericarya and proximal processes of oligodendrocytes in the corpus callosum and internal capsule co-localizing with the myc immunoreactivity ( A and B ) and restricted to CC1 plus; oligodendrocytes in full vector injected mice, while untreated Cx47 knockout mouse brain shows no Cx47 or myc immunoreactivity ( C and F ). Some endogenous green fluorescence is present diffusely in the cytoplasm of all Cx47 knockout oligodendrocytes (including the untreated mice) due to the transgenic expression. Double immunostaining for Cx47 or myc and other cell markers including neuronal NeuN ( G ), astrocytic GFAP ( H ), and microglia Iba1 ( I ) confirms that Cx47 is not expressed in any of these cell types and is restricted to oligodendrocytes. Scale bars thinsp; equals; thinsp;10 thinsp; mu;m. ( J ) Quantification of the percentage of Cx47 expressing oligodendrocytes in the corpus callosum (CC), internal capsule (IC), ventral pons (corticospinal tract area) and cervical spinal cord (SC) anteriolateral white matter of n thinsp; equals; thinsp; 5 mice shows high expression ratios in all areas. ( K ) Counts of Cx47 gap junction plaques per oligodendrocyte confirm the re-establishment of oligodendrocyte gap junctions at levels above those of wild-type mouse brain. ( L ) Immunoblot analysis of brain lysates from three AAV.MBP.Cx47myc injected mice shows the presence of a faint Cx47 band that is absent from non-injected Cx47 knockout mice and is present in a wild-type mouse sample run as positive control (a non-specific band below the specific one is marked with asterisk). Blotting with a myc antibody also shows immunoreactivity in injected mice, which is absent from non-injected knockout and from the wild-type mouse. GAPDH blot is used for loading control. ( M ) Quantification of band intensity shows that Cx47 level in treated knockout mice reached 38.73 thinsp; plusmn; thinsp;11.69 percnt; of the wild-type brain sample. KO thinsp; equals; thinsp;knockout.
    Figure Legend Snippet: AAV.MBP.Cx47myc leads to expression of Cx47 and gap junction formation in oligodendrocytes of Cx47 knockout mice. Double immunostaining for Cx47 (red) and either the myc tag ( A ndash;C ) or the oligodendrocyte marker CC1 ( D ndash;F ) (green) shows Cx47 expression and formation of gap junction (GJ) like plaques (open arrowheads) at the pericarya and proximal processes of oligodendrocytes in the corpus callosum and internal capsule co-localizing with the myc immunoreactivity ( A and B ) and restricted to CC1 plus; oligodendrocytes in full vector injected mice, while untreated Cx47 knockout mouse brain shows no Cx47 or myc immunoreactivity ( C and F ). Some endogenous green fluorescence is present diffusely in the cytoplasm of all Cx47 knockout oligodendrocytes (including the untreated mice) due to the transgenic expression. Double immunostaining for Cx47 or myc and other cell markers including neuronal NeuN ( G ), astrocytic GFAP ( H ), and microglia Iba1 ( I ) confirms that Cx47 is not expressed in any of these cell types and is restricted to oligodendrocytes. Scale bars thinsp; equals; thinsp;10 thinsp; mu;m. ( J ) Quantification of the percentage of Cx47 expressing oligodendrocytes in the corpus callosum (CC), internal capsule (IC), ventral pons (corticospinal tract area) and cervical spinal cord (SC) anteriolateral white matter of n thinsp; equals; thinsp; 5 mice shows high expression ratios in all areas. ( K ) Counts of Cx47 gap junction plaques per oligodendrocyte confirm the re-establishment of oligodendrocyte gap junctions at levels above those of wild-type mouse brain. ( L ) Immunoblot analysis of brain lysates from three AAV.MBP.Cx47myc injected mice shows the presence of a faint Cx47 band that is absent from non-injected Cx47 knockout mice and is present in a wild-type mouse sample run as positive control (a non-specific band below the specific one is marked with asterisk). Blotting with a myc antibody also shows immunoreactivity in injected mice, which is absent from non-injected knockout and from the wild-type mouse. GAPDH blot is used for loading control. ( M ) Quantification of band intensity shows that Cx47 level in treated knockout mice reached 38.73 thinsp; plusmn; thinsp;11.69 percnt; of the wild-type brain sample. KO thinsp; equals; thinsp;knockout.

    Techniques Used: Expressing, Knock-Out, Mouse Assay, Double Immunostaining, Marker, Plasmid Preparation, Injection, Fluorescence, Transgenic Assay, Positive Control

    33) Product Images from "Fractalkine/CX3CR1 Contributes to Endometriosis-Induced Neuropathic Pain and Mechanical Hypersensitivity in Rats"

    Article Title: Fractalkine/CX3CR1 Contributes to Endometriosis-Induced Neuropathic Pain and Mechanical Hypersensitivity in Rats

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2018.00495

    Expressions of fractalkine (FKN) and its receptor CX3CR1 in graft tissue. (A) Immunohistochemical staining for FKN and CX3CR1 in the sciatic nerve of graft tissue. (B) We analyzed FKN and CX3CR1 expression in the sciatic nerve of graft tissue by 1-way ANOVA. (C) Western blotting analysis showed protein levels of membrane-bound FKN, sFKN, and CX3CR1 in graft tissue. (D) Quantification of membrane-bound FKN, sFKN, and CX3CR1 bands in graft tissue, and analysis with 1-way ANOVA. (E) Immunofluorescence staining showed that FKN (red) was mainly expressed in macrophages labeled with Iba1 (green) in graft tissue. (F) The fluorescent images of FKN were statistically graphed for mean density, macrophage-positive cells, DAPI-positive cells, and co-expression in cells under 1-way ANOVA. (G) Immunofluorescent images show that CX3CR1 (red) was highly expressed in nerve fibers as indicated by PGP9.5 (green). The white arrows indicate the co-expression of CX3CR1 and PGP9.5 in cells. (H) Quantitative analysis of CX3CR1-positive cells, PGP9.5-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. (I) CX3CR1 was highly expressed on the myelin sheath when co-stained with MBP (green) using immunohistochemical staining. The white arrows indicate the co-expression of CX3CR1 and MBP in cells. (J) Quantitative analysis of CX3CR1-positive cells, MBP-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. N = 8 rats per group. ** p
    Figure Legend Snippet: Expressions of fractalkine (FKN) and its receptor CX3CR1 in graft tissue. (A) Immunohistochemical staining for FKN and CX3CR1 in the sciatic nerve of graft tissue. (B) We analyzed FKN and CX3CR1 expression in the sciatic nerve of graft tissue by 1-way ANOVA. (C) Western blotting analysis showed protein levels of membrane-bound FKN, sFKN, and CX3CR1 in graft tissue. (D) Quantification of membrane-bound FKN, sFKN, and CX3CR1 bands in graft tissue, and analysis with 1-way ANOVA. (E) Immunofluorescence staining showed that FKN (red) was mainly expressed in macrophages labeled with Iba1 (green) in graft tissue. (F) The fluorescent images of FKN were statistically graphed for mean density, macrophage-positive cells, DAPI-positive cells, and co-expression in cells under 1-way ANOVA. (G) Immunofluorescent images show that CX3CR1 (red) was highly expressed in nerve fibers as indicated by PGP9.5 (green). The white arrows indicate the co-expression of CX3CR1 and PGP9.5 in cells. (H) Quantitative analysis of CX3CR1-positive cells, PGP9.5-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. (I) CX3CR1 was highly expressed on the myelin sheath when co-stained with MBP (green) using immunohistochemical staining. The white arrows indicate the co-expression of CX3CR1 and MBP in cells. (J) Quantitative analysis of CX3CR1-positive cells, MBP-positive cells, DAPI-positive cells, and co-expression in cells using 1-way ANOVA. N = 8 rats per group. ** p

    Techniques Used: Immunohistochemistry, Staining, Expressing, Western Blot, Immunofluorescence, Labeling

    34) Product Images from "Pivotal Role of Brain-Derived Neurotrophic Factor Secreted by Mesenchymal Stem Cells in Severe Intraventricular Hemorrhage in Newborn Rats"

    Article Title: Pivotal Role of Brain-Derived Neurotrophic Factor Secreted by Mesenchymal Stem Cells in Severe Intraventricular Hemorrhage in Newborn Rats

    Journal: Cell Transplantation

    doi: 10.3727/096368916X692861

    Knock down of BDNF in MSCs abolished the therapeutic effects of MSCs in improving brain myelination and in attenuating cell death and reactive gliosis after severe IVH. (A) Representative immunofluorescence photomicrographs of the periventricular area with staining for myelin basic protein (MBP) (green), TUNEL (green), glial fibrillary acidic protein (GFAP) (red), and 4′,6-diamidino-2-pheylindole (DAPI) (blue) (original magnification: 200×, scale bars: 50 μm for MBP and original magnification: 85×, scale bars: 500 μm for TUNEL and GFAP). (B) Western blot assay of MBP, GFAP, and caspase 3 with tubulin in the brain tissue homogenates on P32. Brain MBP expression, cell death, and reactive gliosis evidenced by light intensity of MBP immunofluorescence, average number of TUNEL + cells, and light intensity of GFAP staining, respectively, in the periventricular area (C) and those indicated by the ratio of MBP, caspase 3, and GFAP to tubulin with Western assay, respectively, in the brain tissue homogenates (D) on P32. Data are expressed as mean ± SEM. Abbreviations: NC, normal control rats; IC, IVH control rats; IM, IVH with transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs); IM-cont, IVH with transplantation of scrambled small interfering (si)RNA-transfected hUCB-MSCs; IM-bdnf-kd, transplantation of BDNF siRNA-transfected hUCB-MSCs. * p
    Figure Legend Snippet: Knock down of BDNF in MSCs abolished the therapeutic effects of MSCs in improving brain myelination and in attenuating cell death and reactive gliosis after severe IVH. (A) Representative immunofluorescence photomicrographs of the periventricular area with staining for myelin basic protein (MBP) (green), TUNEL (green), glial fibrillary acidic protein (GFAP) (red), and 4′,6-diamidino-2-pheylindole (DAPI) (blue) (original magnification: 200×, scale bars: 50 μm for MBP and original magnification: 85×, scale bars: 500 μm for TUNEL and GFAP). (B) Western blot assay of MBP, GFAP, and caspase 3 with tubulin in the brain tissue homogenates on P32. Brain MBP expression, cell death, and reactive gliosis evidenced by light intensity of MBP immunofluorescence, average number of TUNEL + cells, and light intensity of GFAP staining, respectively, in the periventricular area (C) and those indicated by the ratio of MBP, caspase 3, and GFAP to tubulin with Western assay, respectively, in the brain tissue homogenates (D) on P32. Data are expressed as mean ± SEM. Abbreviations: NC, normal control rats; IC, IVH control rats; IM, IVH with transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs); IM-cont, IVH with transplantation of scrambled small interfering (si)RNA-transfected hUCB-MSCs; IM-bdnf-kd, transplantation of BDNF siRNA-transfected hUCB-MSCs. * p

    Techniques Used: Immunofluorescence, Staining, TUNEL Assay, Western Blot, Expressing, Transplantation Assay, Derivative Assay, Transfection

    35) Product Images from "Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells"

    Article Title: Rapid functional genetics of the oligodendrocyte lineage using pluripotent stem cells

    Journal: Nature Communications

    doi: 10.1038/s41467-018-06102-7

    Cellular profiling of spontaneous and purposely generated mutant oligodendrocyte alleles. a Diagram indicating that shiverer mice harbor a ~20-kilobase (kb) homozygous deletion encompassing exons 2–7 of the MBP gene. A Sanger sequencing trace shows the breakpoint of the shiverer deletion. b Diagram indicating the location of the two gRNAs designed to target MYRF . A Sanger sequencing trace shows the location of the homozygous deletion of exon 1. c Quantification of transcription factors Olig2, Nkx2.2, and Sox10 at passage 3 of the differentiation protocol. n = 3 shiverer cell lines; n = 3 replicate wells per cell line; > 179,500 cells scored per well. n = 3 MYRF KO and wild-type (WT) mESC replicate wells per cell line; > 700 cells scored per well. Data are represented as means ± SEM. d Fluorescent images of WT iPSC, shiverer , and MYRF KO OPCs expressing canonical OPC markers Olig2, Nkx2.2, and Sox10. Scale bar, 50 µm. e Cell surface immunostaining of the immature oligodendrocyte marker O4, after treatment with T3, of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. f Representative images of differentiated OPCs immunostained for mature oligodendrocyte markers MBP and PLP1, 72 h post treatment with T3 of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. g Representative images of OPC/DRG co-cultures stained for MBP and neurofilament (NF) at day 10 from WT iPSC, shiverer , and MYRF KO OPCs stained for PLP1 or MBP after being co-cultured for 10 days with NF+ embryonic rat DRGs. Scale bar, 50 µm
    Figure Legend Snippet: Cellular profiling of spontaneous and purposely generated mutant oligodendrocyte alleles. a Diagram indicating that shiverer mice harbor a ~20-kilobase (kb) homozygous deletion encompassing exons 2–7 of the MBP gene. A Sanger sequencing trace shows the breakpoint of the shiverer deletion. b Diagram indicating the location of the two gRNAs designed to target MYRF . A Sanger sequencing trace shows the location of the homozygous deletion of exon 1. c Quantification of transcription factors Olig2, Nkx2.2, and Sox10 at passage 3 of the differentiation protocol. n = 3 shiverer cell lines; n = 3 replicate wells per cell line; > 179,500 cells scored per well. n = 3 MYRF KO and wild-type (WT) mESC replicate wells per cell line; > 700 cells scored per well. Data are represented as means ± SEM. d Fluorescent images of WT iPSC, shiverer , and MYRF KO OPCs expressing canonical OPC markers Olig2, Nkx2.2, and Sox10. Scale bar, 50 µm. e Cell surface immunostaining of the immature oligodendrocyte marker O4, after treatment with T3, of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. f Representative images of differentiated OPCs immunostained for mature oligodendrocyte markers MBP and PLP1, 72 h post treatment with T3 of WT iPSC, shiverer , and MYRF KO OPCs. Scale bar, 50 µm. g Representative images of OPC/DRG co-cultures stained for MBP and neurofilament (NF) at day 10 from WT iPSC, shiverer , and MYRF KO OPCs stained for PLP1 or MBP after being co-cultured for 10 days with NF+ embryonic rat DRGs. Scale bar, 50 µm

    Techniques Used: Generated, Mutagenesis, Mouse Assay, Sequencing, Expressing, Immunostaining, Marker, Staining, Cell Culture

    36) Product Images from "Neurons define non-myelinated axon segments by the regulation of galectin-4-containing axon membrane domains"

    Article Title: Neurons define non-myelinated axon segments by the regulation of galectin-4-containing axon membrane domains

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-12295-6

    Axonal G4Ds are not myelinated in neuron/oligodendrocyte co-cultures. ( a ) Hippocampal neurons (stage 3; 72 hiv ) were immunolabelled for Gal-4 (red) without permeabilization. Gal-4 is detected in segments (arrowheads) along the axonal plasma membrane. ( b ) HA-tagged Gal-4 (Gal-4-HA) transfected hippocampal neurons were immunolabelled for HA (red) without permeabilization. Gal-4-HA is also detected in axon membrane segments (arrowheads). ( c ) Gal-4-HA transfected astrocytes were immunolabelled for HA (red) without permeabilization. Expressed Gal-4-HA forms “patches” on the cell surface (Gal-4-containing membrane domains; arrows). ( d – g ) Hippocampal neurons and oligodendrocytes were co-cultured for different time periods. The ICC for Gal-4 (red) was performed under non-permeabilizing conditions to detect Gal-4 associated to axon surface. Cells were then permeabilized and immunolabelled for acetylated tubulin (blue) and MBP (green) to label axons and myelin, respectively. ( d ) The amount of Gal-4 associated to the axon membrane is not affected by the contact with oligodendrocytes at early stage of differentiation (arrowheads). ( e ) Axon points where initial contacts of myelinating oligodendrocytes occur are devoid of membrane Gal-4 (arrowheads). ( f ) At more advanced stages of myelination in vitro , membrane Gal-4 is absent in myelinated axon segments (arrowheads). A 3D reconstruction from a stack of Z-planes processed to simulate opaque surfaces show that the myelin sheet (green) entirely wraps the portion of the axon devoid of Gal-4 (red) as seen from the top and bottom surfaces of the reconstructed image. ( g ) Membrane Gal-4 and MBP expression along the axon (dotted line in MBP picture depicts an example of axonal “region of interest” ROI), measured as percentages of the maximum fluorescence intensity in each channel, is plotted in the line graph (MBP green line, Gal-4 red line). Bar graph shows that average Gal-4 expression in non-myelinated axon segments (black bar) is significantly higher than that of myelinated ones (gray bar). Axon segments are considered “myelinated” when measured MBP fluorescence intensity is over 60% of its maximum, and “non-myelinated” when it is lower than 30% of the maximum (values are means + s.e.m. of 3 experiments, n = 38 and n = 35, respectively; *p
    Figure Legend Snippet: Axonal G4Ds are not myelinated in neuron/oligodendrocyte co-cultures. ( a ) Hippocampal neurons (stage 3; 72 hiv ) were immunolabelled for Gal-4 (red) without permeabilization. Gal-4 is detected in segments (arrowheads) along the axonal plasma membrane. ( b ) HA-tagged Gal-4 (Gal-4-HA) transfected hippocampal neurons were immunolabelled for HA (red) without permeabilization. Gal-4-HA is also detected in axon membrane segments (arrowheads). ( c ) Gal-4-HA transfected astrocytes were immunolabelled for HA (red) without permeabilization. Expressed Gal-4-HA forms “patches” on the cell surface (Gal-4-containing membrane domains; arrows). ( d – g ) Hippocampal neurons and oligodendrocytes were co-cultured for different time periods. The ICC for Gal-4 (red) was performed under non-permeabilizing conditions to detect Gal-4 associated to axon surface. Cells were then permeabilized and immunolabelled for acetylated tubulin (blue) and MBP (green) to label axons and myelin, respectively. ( d ) The amount of Gal-4 associated to the axon membrane is not affected by the contact with oligodendrocytes at early stage of differentiation (arrowheads). ( e ) Axon points where initial contacts of myelinating oligodendrocytes occur are devoid of membrane Gal-4 (arrowheads). ( f ) At more advanced stages of myelination in vitro , membrane Gal-4 is absent in myelinated axon segments (arrowheads). A 3D reconstruction from a stack of Z-planes processed to simulate opaque surfaces show that the myelin sheet (green) entirely wraps the portion of the axon devoid of Gal-4 (red) as seen from the top and bottom surfaces of the reconstructed image. ( g ) Membrane Gal-4 and MBP expression along the axon (dotted line in MBP picture depicts an example of axonal “region of interest” ROI), measured as percentages of the maximum fluorescence intensity in each channel, is plotted in the line graph (MBP green line, Gal-4 red line). Bar graph shows that average Gal-4 expression in non-myelinated axon segments (black bar) is significantly higher than that of myelinated ones (gray bar). Axon segments are considered “myelinated” when measured MBP fluorescence intensity is over 60% of its maximum, and “non-myelinated” when it is lower than 30% of the maximum (values are means + s.e.m. of 3 experiments, n = 38 and n = 35, respectively; *p

    Techniques Used: Transfection, Cell Culture, Immunocytochemistry, In Vitro, Expressing, Fluorescence

    Related Articles

    Incubation:

    Article Title: Combined Adenovirus-Mediated Artificial microRNAs Targeting mfgl2, mFas, and mTNFR1 Protect against Fulminant Hepatic Failure in Mice
    Article Snippet: Immunohistochemical staining Liver tissues were isolated, sliced, fixed, dehydrated, and then subjected to immunohistochemistry using PV6001 PowerVision Two-Step Histostaining Reagent (ZSGB-BIO, Beijing, China) according to the manufacturer’s instructions. .. Liver tissue sections were incubated with a rabbit polyclonal antibody against mfgl2 prothrombinase (1:200 dilution; Abcam, Cambridge, UK), mFas (1:50 dilution; Abcam, Cambridge, UK), and mTNFR1 (1:1000 dilution; Abcam, Cambridge, UK) or with a polyclonal antibody against fibrinogen (1:1000 dilution, Dako, Glostrup, Denmark) as previously described [ ]. .. After incubation with horseradish peroxidase (HRP)-labeled goat IgG fraction to rabbit IgG Fc, the target protein was detected using a diaminobenzidine (DAB) kit (ZSGB-BIO, Beijing, China).

    Article Title: Biological Characterization of Gene Response to Insulin-Induced Hypoglycemia in Mouse Retina
    Article Snippet: .. Briefly, frozen retina sections were blocked in PBS with 3% normal goat serum (Sigma, Buchs, Switzerland) and 0.2% Triton X-100 (Sigma) for 1 h at RT and then incubated with antibodies against cleaved Caspase3 (dilution 1/500, Cell Signaling Technology, Inc. Danvers, MA, USA), GPX3 (dilution 1/50, Abcam, Cambridge, MA, USA), GSTO-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA), FIBULIN-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA) and VERSICAN (dilution 1/100, Abcam, Cambridge, MA, USA) in the blocking buffer overnight at 4°C. .. Thereafter, sections were incubated again in blocking buffer for 30 min at RT before being incubated for 1 h at RT with FITC Alexa-Fluor 594 goat anti-rabbit antibody or with FITC Alexa-Fluor 594 sheep anti-goat antibody (dilution 1/2’000,) depending of primary antibody used.

    other:

    Article Title: Roles of Heparan Sulfate Sulfation in Dentinogenesis *
    Article Snippet: To investigate the affinity of Wnt10a toward HS, which had been either pretreated with active Sulf1 (termed HS-6S) or not (HS+6S), we used columns with immobilized HS-6S and HS+6S.

    Article Title: Transcriptomic Landscape of von Economo Neurons in Human Anterior Cingulate Cortex Revealed by Microdissected-Cell RNA Sequencing
    Article Snippet: The IF images showed that all four genes were predominantly expressed in VENs with three of them (LYPD1 , SULF2, and CHST8 ) showing strong nuclear signals (Fig. A–P ).

    Blocking Assay:

    Article Title: Biological Characterization of Gene Response to Insulin-Induced Hypoglycemia in Mouse Retina
    Article Snippet: .. Briefly, frozen retina sections were blocked in PBS with 3% normal goat serum (Sigma, Buchs, Switzerland) and 0.2% Triton X-100 (Sigma) for 1 h at RT and then incubated with antibodies against cleaved Caspase3 (dilution 1/500, Cell Signaling Technology, Inc. Danvers, MA, USA), GPX3 (dilution 1/50, Abcam, Cambridge, MA, USA), GSTO-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA), FIBULIN-1 (dilution 1/200, Santa-Cruz Biotechnology, Dallas, TX, USA) and VERSICAN (dilution 1/100, Abcam, Cambridge, MA, USA) in the blocking buffer overnight at 4°C. .. Thereafter, sections were incubated again in blocking buffer for 30 min at RT before being incubated for 1 h at RT with FITC Alexa-Fluor 594 goat anti-rabbit antibody or with FITC Alexa-Fluor 594 sheep anti-goat antibody (dilution 1/2’000,) depending of primary antibody used.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Abcam anti mbp
    Loss of HMGNs affects oligodendrocyte development in mouse spinal cord. ( A–D ) Immunofluorescence showing decreased OLIG2 positive cells in both white matter ( A ) and gray matter ( C ) of DKO mouse spinal cords. ( B, D) Quantifications of four WT and four DKO mice. For each mouse, > 1500 cells from six (white matter) or four (gray matter) different regions of spinal cord sections were counted. ( E ) Immunofluorescence showing decreased <t>MBP</t> level in DKO spinal cords. ( F ) Quantification of panel J, eight WT and eight DKO images were quantified by ImageJ. ( G ) Western blots showing the expression of oligodendrocyte markers in WT and DKO mouse spinal cords. ( H ) Quantifications of panel L, expression levels normalized to <t>β-actin.</t> All the data are presented as mean ± SEM (* P
    Anti Mbp, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti mbp/product/Abcam
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti mbp - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

    mbp  (Abcam)
    94
    Abcam mbp
    Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into <t>MBP</t> Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human <t>GFAP</t> (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p
    Mbp, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mbp/product/Abcam
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mbp - by Bioz Stars, 2021-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Loss of HMGNs affects oligodendrocyte development in mouse spinal cord. ( A–D ) Immunofluorescence showing decreased OLIG2 positive cells in both white matter ( A ) and gray matter ( C ) of DKO mouse spinal cords. ( B, D) Quantifications of four WT and four DKO mice. For each mouse, > 1500 cells from six (white matter) or four (gray matter) different regions of spinal cord sections were counted. ( E ) Immunofluorescence showing decreased MBP level in DKO spinal cords. ( F ) Quantification of panel J, eight WT and eight DKO images were quantified by ImageJ. ( G ) Western blots showing the expression of oligodendrocyte markers in WT and DKO mouse spinal cords. ( H ) Quantifications of panel L, expression levels normalized to β-actin. All the data are presented as mean ± SEM (* P

    Journal: Nucleic Acids Research

    Article Title: Interplay between H1 and HMGN epigenetically regulates OLIG1 2 expression and oligodendrocyte differentiation

    doi: 10.1093/nar/gkw1222

    Figure Lengend Snippet: Loss of HMGNs affects oligodendrocyte development in mouse spinal cord. ( A–D ) Immunofluorescence showing decreased OLIG2 positive cells in both white matter ( A ) and gray matter ( C ) of DKO mouse spinal cords. ( B, D) Quantifications of four WT and four DKO mice. For each mouse, > 1500 cells from six (white matter) or four (gray matter) different regions of spinal cord sections were counted. ( E ) Immunofluorescence showing decreased MBP level in DKO spinal cords. ( F ) Quantification of panel J, eight WT and eight DKO images were quantified by ImageJ. ( G ) Western blots showing the expression of oligodendrocyte markers in WT and DKO mouse spinal cords. ( H ) Quantifications of panel L, expression levels normalized to β-actin. All the data are presented as mean ± SEM (* P

    Article Snippet: Anti-H3K27me3, anti-EZH2, anti-H3, anti-PGDFR-α, anti-CNPase, anti-PLP, anti-MBP and anti-β-actin antibodies were purchased from Abcam.

    Techniques: Immunofluorescence, Mouse Assay, Western Blot, Expressing

    Combination treatment with Sephin1 and IFN-β alleviates and delays clinical symptoms of EAE as well as reduces the oligodendrocytes and myelin loss during EAE course. ( A ) Clinical scores of C57BL/6 female mice immunized with MOG 35-55 /CFA to induce chronic EAE, treated with vehicle ( n = 12), 5000 U of IFN-β ( n = 12), 8 mg/kg of Sephin1 or Sephin1 combined with 5000 U of IFNβ ( n = 10) daily from PID 7 to the end of the study. ( B ) Average onset of disease, peak of disease and average peak score of all treatment groups. ( C and D ) Immunofluorescent staining for TPPP (green) and MBP (red) of lumbar spinal cord sections from PID 17 ( C ) and PID 30 ( D ). Scale bar = 100 μm. ( E ) Quantification of cells positive for TPPP in the lesion areas. ( F ) Average percentage of demyelinated areas/white matter areas measured from MBP staining. * P

    Journal: Brain

    Article Title: Sephin1, which prolongs the integrated stress response, is a promising therapeutic for multiple sclerosis

    doi: 10.1093/brain/awy322

    Figure Lengend Snippet: Combination treatment with Sephin1 and IFN-β alleviates and delays clinical symptoms of EAE as well as reduces the oligodendrocytes and myelin loss during EAE course. ( A ) Clinical scores of C57BL/6 female mice immunized with MOG 35-55 /CFA to induce chronic EAE, treated with vehicle ( n = 12), 5000 U of IFN-β ( n = 12), 8 mg/kg of Sephin1 or Sephin1 combined with 5000 U of IFNβ ( n = 10) daily from PID 7 to the end of the study. ( B ) Average onset of disease, peak of disease and average peak score of all treatment groups. ( C and D ) Immunofluorescent staining for TPPP (green) and MBP (red) of lumbar spinal cord sections from PID 17 ( C ) and PID 30 ( D ). Scale bar = 100 μm. ( E ) Quantification of cells positive for TPPP in the lesion areas. ( F ) Average percentage of demyelinated areas/white matter areas measured from MBP staining. * P

    Article Snippet: Primary antibodies include the following: anti-MBP (Abcam, ab24567, 1:700), anti-TPPP (Thermo Fisher Scientific, PA5-19243, 1:100), anti-p-eIF2α (Abcam, ab32157, 1:100; Thermo Fisher Scientific, MA5-15133, 1:50), anti-CD3 (Santa Cruz, sc-18843, 1:200), anti-CD11b (Bio-Rad, MCA711, 1:50), anti-NG2 (Millipore, Ab5320, 1:100), anti-neurofilament (Millipore, AB5539, 1:500), anti-GFAP (Millipore, AB5541, 1:200), anti-caspase 3 (Abcam, AB2302, 1:50), anti-Ki67 (Abcam, AB15580, 1:100), anti-PDGFR-alpha (BD Biosciences, 558774, 1:100), and anti-SOX10 (R & D, AF2864, 1:100).

    Techniques: Mouse Assay, Staining

    AATYK knockdown does not affect the survival but maintains proliferation of OPCs. A Ki67 immunostaining showed there were more Ki67 + and GFP + co-labeled cells in the sh- AATYK -treated OPCs than in the control. B Percentages of Ki67 + and GFP + co-labelled OLs in GFP + cells among RNAi- AATYK cells were significantly higher than in the control. C – D Compared to control cells, the expression of MBP protein was significantly decreased in shRNA- AATYK -treated cells and there was no difference in caspase3 between the two groups. Scale bar, 100 μm.

    Journal: Neuroscience Bulletin

    Article Title: AATYK is a Novel Regulator of Oligodendrocyte Differentiation and Myelination

    doi: 10.1007/s12264-018-0218-6

    Figure Lengend Snippet: AATYK knockdown does not affect the survival but maintains proliferation of OPCs. A Ki67 immunostaining showed there were more Ki67 + and GFP + co-labeled cells in the sh- AATYK -treated OPCs than in the control. B Percentages of Ki67 + and GFP + co-labelled OLs in GFP + cells among RNAi- AATYK cells were significantly higher than in the control. C – D Compared to control cells, the expression of MBP protein was significantly decreased in shRNA- AATYK -treated cells and there was no difference in caspase3 between the two groups. Scale bar, 100 μm.

    Article Snippet: SDS-PAGE gels were subsequently detected with anti-MBP (Abcam, 1:2 000), anti-caspase3 (Millipore, 1:1 000), and anti-GAPDH (Sigma, 1:5 000) antibodies.

    Techniques: Immunostaining, Labeling, Expressing, shRNA

    Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into MBP Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human GFAP (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p

    Journal: bioRxiv

    Article Title: Familial t(1;11) translocation is associated with disruption of white matter structural integrity and oligodendrocyte-myelin dysfunction

    doi: 10.1101/657163

    Figure Lengend Snippet: Case and control OPCs transplanted neonatally in the hypomyelinated show reduction in myelin sheath length at 8- and 13-weeks post-transplantation (a,b) Human OPCs transplanted into MBP Shi/Shi ;Rag2 -/- neonates (at P1-P2) efficiently engraft and differentiate to oligodendrocytes (arrowhead in a) and astrocytes (arrow in b) in the white matter (dotted lines) by 8 weeks. Human cells are identified by immunoreactivity against human nuclei (red) and MBP (green) in (a) and with human GFAP (red) and MBP (green) in (b). (c-f) Individual human oligodendrocytes from Control1 and 2 (c,d) and Case2, 4 lines (e.f) were imaged and length of myelin segments measured. Arrowheads demarcate the beginning and end of individual myelin segments visualized in 3D. (g-h) Quantification of average myelin sheath lengths from (c-f) shows a severe reduction in cases at 8 weeks (g) and 13 weeks post transplantation (h). Mean with upper and lower quartiles shown. Scatter points represent outliers. p

    Article Snippet: Antibodies; OLIG2 (1:200, Millipore), PDGFRα (1:500, Cell Signalling), O4 (1:500, R & D Systems), MBP (1:50, Abcam), GFAP (1:500, DAKO), GFAP (1:500, Cy3 conjugated; Sigma), CASPR (1:1000, Abcam), Sox2 (1:1000, Abcam), Oct3/4 (1:250, Santa Cruz), Nanog (1:100, R & D Systems), TRA1-60 (1:100, StemGent), Human Nuclei (1:300, Millipore), Human GFAP (1:1000, Biolegend), Neurofilament-H (1:10,000, Biolegend).

    Techniques: Transplantation Assay

    Efficient conversion of control and case iPS cells to oligodendroglial lineage (a1-a7) Both control and case iPS lines could be patterned to OLIG2+ precursor cells under conditions described in Methods. (b1-b7) Further differentiation of cells from (a) gave PDFGRα+ OPCs under proliferative conditions containing FGF2 and PDGF-AA (c1-c7) Removal of mitogens produced mature oligodendrocytes that co-stained for O4 and MBP. (d-f) Quantification of OLIG2+ cells on day1 (d) PDFGRα+ OPCs on day7 (e) and O4+ oligodendrocytes at day21 (f) shows a significant increase in oligodendrocytes in case lines (n= 3 independent conversions for each line, unpaired t-test used to analyze p-values) Scale: (a-c) 50 μm

    Journal: bioRxiv

    Article Title: Familial t(1;11) translocation is associated with disruption of white matter structural integrity and oligodendrocyte-myelin dysfunction

    doi: 10.1101/657163

    Figure Lengend Snippet: Efficient conversion of control and case iPS cells to oligodendroglial lineage (a1-a7) Both control and case iPS lines could be patterned to OLIG2+ precursor cells under conditions described in Methods. (b1-b7) Further differentiation of cells from (a) gave PDFGRα+ OPCs under proliferative conditions containing FGF2 and PDGF-AA (c1-c7) Removal of mitogens produced mature oligodendrocytes that co-stained for O4 and MBP. (d-f) Quantification of OLIG2+ cells on day1 (d) PDFGRα+ OPCs on day7 (e) and O4+ oligodendrocytes at day21 (f) shows a significant increase in oligodendrocytes in case lines (n= 3 independent conversions for each line, unpaired t-test used to analyze p-values) Scale: (a-c) 50 μm

    Article Snippet: Antibodies; OLIG2 (1:200, Millipore), PDGFRα (1:500, Cell Signalling), O4 (1:500, R & D Systems), MBP (1:50, Abcam), GFAP (1:500, DAKO), GFAP (1:500, Cy3 conjugated; Sigma), CASPR (1:1000, Abcam), Sox2 (1:1000, Abcam), Oct3/4 (1:250, Santa Cruz), Nanog (1:100, R & D Systems), TRA1-60 (1:100, StemGent), Human Nuclei (1:300, Millipore), Human GFAP (1:1000, Biolegend), Neurofilament-H (1:10,000, Biolegend).

    Techniques: Produced, Staining