rabbit anti olig1  (PhosphoSolutions)

Journal: PLoS ONE

Article Title: Neuroprotective Effect of Transplanted Human Embryonic Stem Cell-Derived Neural Precursors in an Animal Model of Multiple Sclerosis

doi: 10.1371/journal.pone.0003145

Figure Lengend Snippet: (A–H) Immunofluorescence stainings of brain sections demonstrating the migration and differentiation of transplanted NPs, which were identified by the expression of human mitochondria (A, C–G), human nuclear antigens (B) and GFP (inset in A, H). (A): The NPs migrated extensively into white matter areas of the CNS such as the corpus callosum ( CC ) and were not observed in grey areas such as subcortical grey matter ( SGM ). Co-staining against the oligodendroglial marker, O4 (red) was used to identify the white matter. (B–H): Most of the transplanted cells either remained as uncommitted NPs expressing Musashi (B) or differentiated into early neuronal/oligodendroglial progenitors expressing Olig1 (C) or Olig2 (D). Further differentiation into more committed neuronal progenitors, oligodendrocyte progenitors or astrocytes expressing NGN2 (E), NG2 (F) and GFAP (G), respectively, was infrequent (∼1% of the transplanted cells per each of the cell types). Terminal differentiation of the transplanted NPs into GalC-expressing mature oligodendrocytes (H) was rare (<0.01% of the transplanted cells). Nuclei in A–H are counterstained with DAPI (blue). I–J: Toluidine blue stained transverse semi-thin sections of resin embedded spinal cords of NP-transplanted (J) and control (I) animals. In NP-transplanted animals, there were less demyelinated (arrows in I) and more normally myelinated axons than in controls. In both groups there were very rare remyelinated axons (G ratio>0.8). Scale bars: 15 µm (A–H), 5 µm (I–J).

Article Snippet: Immunofluorescent staining of NPs in vitro and in vivo: The following primary antibodies were used: Rabbit IgG anti-GFP (1∶100, Chemicon), mouse IgG anti-human specific mitochondria (1∶200, Chemicon), mouse IgM anti-A2B5 (1∶1, ATCC), mouse IgM anti-PSA-NCAM (1∶200, Chemicon), rabbit IgG anti-nestin (1∶50, Chemicon), rabbit anti-musashi (1∶100, Chemicon), mouse IgG anti-human nuclei (1∶50, Chemicon), rabbit IgG anti-NG2 (1∶50, Chemicon), mouse IgM anti-PDGFRα (1∶20, R&D), rabbit IgG anti-NGN2 (1∶300, Chemicon), rabbit anti-GalC (1∶20, Chemicon), mouse IgM anti-O4 (1∶20, Chemicon), rabbit IgG anti-MAP2 (1∶200, Chemicon), mouse IgG anti-β tubulin III (1∶2000, Sigma), rabbit anti-GFAP (1∶100, Dako), rabbit IgG anti-olig1 (1∶20, Chemicon) and goat anti-olig2 (1∶30, R&D).

Techniques: Immunofluorescence, Migration, Expressing, Staining, Marker

Journal: bioRxiv

Article Title: Region-specific, maladaptive, gray matter myelination is associated with differential susceptibility to stress-induced behavior in rats and humans

doi: 10.1101/2021.02.15.431176

Figure Lengend Snippet: (A) Injection schematic and lentiviral constructs. CMV, human cytomegalovirus promoter; EGFP, enhanced green fluorescent protein; WPRE, Woodchuck hepatitis virus posttranscriptional regulatory element; Olig1, oligodendrocyte transcription factor 1. (B) Experimental timeline. BrdU, bromodeoxyuridine; LD, light- dark box. (C) Representative image of viral spread through the dorsal dentate gyrus. Lenti, lentivirus; DAPI, 4′,6-diamidino-2-phenylindole; GFP, green fluorescent protein. Scale bar = 200 μm. (D) Representative images of GFP and Olig1 immunohistochemistry 7 weeks post-injection. Scale bar = 50 μm. (E) Dentate gyrus Olig1 cell densities (GFP: 28.11 ± 2.33 cells/mm 2 , Olig1: 41.54 ± 3.72 cells/mm 2 ; two-tailed t test: t(10) = 3.06, p = 0.012; n = 6 per group). (F) Composite behavior scores (GFP: 4.4 ± 0.9, Olig1: 9.7 ± 2.0; two-tailed t test: t(14) = 2.58, p = 0.022; nGFP = 9, nOlig1 = 7). (G) Composite behavior scores from avoidance tests (GFP: 3.3 ± 0.7, Olig1: 7.0 ± 1.6; two-tailed t test: t(14) = 2.21, p = 0.044; nGFP = 9, nOlig1 = 7). (H) Pearson correlation of Olig1 cell densities to avoidance composite scores from Lenti-Olig1 animals (Pearson correlation: r = 0.86, p = 0.028; n = 6). (I) Composite behavior scores from the acoustic startle response (ASR) test (GFP: 1.1 ± 0.5, Olig1: 2.7 ± 0.5; two-tailed t test: t2,14 = 2.24, p = 0.042; nGFP = 9, nOlig1 = 7). (J) Pearson correlation of Olig1 cell densities to composite startle scores from Lenti-Olig1 animals (Pearson correlation: r = 0.23, p = 0.66; n = 6). For all plots, individual data points are shown with mean ± SEM, * p < 0.05.

Article Snippet: Primary antibodies were rat anti-myelin basic protein (MBP; 1:500; Abcam, Cambridge, UK), rabbit anti-glutathione S-transferase π (GSTπ; 1:1000; MBL International, Woburn, MA), sheep anti-carbonic anhydrase II (CAII; 1:10000; AbD Serotec, Hercules, CA), rabbit anti-Olig1 (1:1000; MilliporeSigma, Burlington, MA), and chicken anti-green fluorescent protein (GFP; 1:750; Abcam, Cambridge, UK).

Techniques: Injection, Construct, Immunohistochemistry, Two Tailed Test

Journal: Cell stem cell

Article Title: OLIG2 Drives Abnormal Neurodevelopmental Phenotypes in Human iPSC-Based Organoid and Chimeric Mouse Models of Down Syndrome

doi: 10.1016/j.stem.2019.04.014

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-OLIG1 , Phosphosolutions , Cat# 1537; RRID: AB_2492192.

Techniques: Virus, shRNA, Recombinant, Gene Expression, Fractionation, Luciferase, Software

Journal: Brain Pathology

Article Title: Complement C3a induces axonal hypomyelination in the periventricular white matter through activation of WNT/β‐catenin signal pathway in septic neonatal rats experimentally induced by lipopolysaccharide

doi: 10.1111/bpa.12798

Figure Lengend Snippet: Primary antibodies used in experiments.

Article Snippet: Image J software was used to quantify the intensity of the protein band (n = 6 each). table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Antibody Host Company Cat. No. RRID Application (concentration) C3a/C3 Chicken Abcam ab48581 AB_869211 WB (1:1000)/IF (1:200) C3aR Rabbit Abcam ab140777 AB_2687440 IF (1:100) C3aR Mouse Santa Cruz Biotechnology sc‐133172 AB_2066736 WB (1:1000)/IF (1:50) TMEM119 Rabbit Abcam ab209064 AB_2800343 IF (1:100) GFAP Mouse Abcam ab4648 AB_449329 IF (1:200) CNPase Mouse Cell Signaling Technology 5664 AB_10705455 WB (1:1000)/IF (1:200) MBP Rabbit Abcam ab62631 AB_956157 WB (1:1000)/IF (1:200) PLP Rabbit Abcam ab28486 AB_776593 WB (1:1000) MAG Mouse Abcam ab89780 AB_2042411 WB (1:1000) Olig1 Rabbit Abcam ab124908 AB_10972689 WB (1:1000) Olig2 Rabbit Abcam ab109186 AB_10861310 WB (1:1000)/IF (1:200) SOX10 Rabbit Abcam ab155279 AB_2650603 WB (1:1000) NG2 Mouse Abcam ab50009 AB_881569 WB (1:1000)/IF (1:200) CC1 Mouse Abcam ab16794 AB_443473 WB (1:1000)/IF (1:200) Ki‐67 Rabbit Cell Signaling Technology 9129 AB_2687446 IF (1:200) β‐catenin Rabbit Abcam ab32572 AB_725966 WB (1:1000) p‐β‐catenin Rabbit Cell Signaling Technology 9561 AB_331729 WB (1:1000) Tcf4 Mouse Novus H00006925‐M03 AB_2199267 WB (1:1000) Axin2 Rabbit Abcam ab32197 AB_2290204 WB (1:1000) β‐actin Mouse Cell Signaling Technology 58169 AB_2750839 WB (1:3000) GAPDH Rabbit Cell Signaling Technology 3683 AB_1642205 WB (1:3000) Open in a separate window Primary antibodies used in experiments.

Techniques: Concentration Assay

Journal: Brain Pathology

Article Title: Complement C3a induces axonal hypomyelination in the periventricular white matter through activation of WNT/β‐catenin signal pathway in septic neonatal rats experimentally induced by lipopolysaccharide

doi: 10.1111/bpa.12798

Figure Lengend Snippet: C3aRa administration upregulated the expression of transcription factors involved in oligodendrocyte maturation and differentiation. Immunofluorescence staining showing Olig2 immunoreactive oligodendrocytes (red) and DAPI (blue) in the PWM in postnatal rats at 7, 14 and 28 days after LPS injection (B, E, H) or LPS + C3aRa administration (C, F, I) and their matching controls (A, D, G) at the magnification of ×40 (n = 5). Panel J shows Olig1 (26 kDa), Olig2 (37 kDa), SOX10 (49 kDa) and β‐actin (42 kDa) immunoreactive bands (n = 6). (K) Bar graph shows the number of Olig2+/DAPI+ in the PWM at 7, 14 and 28 days after LPS injection after LPS injection, LPS + C3aRa administration and their matching controls (n = 5). (L–N) Bar graphs depict the optical density of Olig1, Olig2 and SOX10 expression shown in J (n = 6). For statistical analysis, two‐way ANOVA with Tukey's post hoc test was used for K, L, M and N and presented as the mean ± standard error of measurement (SEM). Scale bars: 20 μm. *P < 0.05, **P < 0.01.

Article Snippet: Image J software was used to quantify the intensity of the protein band (n = 6 each). table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Antibody Host Company Cat. No. RRID Application (concentration) C3a/C3 Chicken Abcam ab48581 AB_869211 WB (1:1000)/IF (1:200) C3aR Rabbit Abcam ab140777 AB_2687440 IF (1:100) C3aR Mouse Santa Cruz Biotechnology sc‐133172 AB_2066736 WB (1:1000)/IF (1:50) TMEM119 Rabbit Abcam ab209064 AB_2800343 IF (1:100) GFAP Mouse Abcam ab4648 AB_449329 IF (1:200) CNPase Mouse Cell Signaling Technology 5664 AB_10705455 WB (1:1000)/IF (1:200) MBP Rabbit Abcam ab62631 AB_956157 WB (1:1000)/IF (1:200) PLP Rabbit Abcam ab28486 AB_776593 WB (1:1000) MAG Mouse Abcam ab89780 AB_2042411 WB (1:1000) Olig1 Rabbit Abcam ab124908 AB_10972689 WB (1:1000) Olig2 Rabbit Abcam ab109186 AB_10861310 WB (1:1000)/IF (1:200) SOX10 Rabbit Abcam ab155279 AB_2650603 WB (1:1000) NG2 Mouse Abcam ab50009 AB_881569 WB (1:1000)/IF (1:200) CC1 Mouse Abcam ab16794 AB_443473 WB (1:1000)/IF (1:200) Ki‐67 Rabbit Cell Signaling Technology 9129 AB_2687446 IF (1:200) β‐catenin Rabbit Abcam ab32572 AB_725966 WB (1:1000) p‐β‐catenin Rabbit Cell Signaling Technology 9561 AB_331729 WB (1:1000) Tcf4 Mouse Novus H00006925‐M03 AB_2199267 WB (1:1000) Axin2 Rabbit Abcam ab32197 AB_2290204 WB (1:1000) β‐actin Mouse Cell Signaling Technology 58169 AB_2750839 WB (1:3000) GAPDH Rabbit Cell Signaling Technology 3683 AB_1642205 WB (1:3000) Open in a separate window Primary antibodies used in experiments.

Techniques: Expressing, Immunofluorescence, Staining, Injection

Journal: Brain Pathology

Article Title: Complement C3a induces axonal hypomyelination in the periventricular white matter through activation of WNT/β‐catenin signal pathway in septic neonatal rats experimentally induced by lipopolysaccharide

doi: 10.1111/bpa.12798

Figure Lengend Snippet: C3a inhibits the differentiation and maturation of OPCs in vitro. Immunofluorescence images of cultured OPCs showing the expression of MBP (A–D, green), NG2 (E–H, green) and DAPI (blue) at 4 days after the C3a, C3a + C3aRa and C3aRa treatment when compared with the corresponding control at the magnification of ×40 (n = 5). Panel I shows the immunoreactive bands of PLP (26 kDa), MBP (33 kDa), CNPase (47 kDa), MAG (69 kDa), NG2 (251 kDa), CC1 (310 kDa), Olig1 (26 kDa), Olig2 (37 kDa), SOX10 (49 kDa) and β‐actin (42 kDa) after C3a administration or C3a + C3aRa treatment or C3aRa treatment and the corresponding control (n = 6). Bar graphs (J–R) show the optical density of protein expression shown in I (n = 6). (S, T) Bar graphs show the percentage of MBP+/DAPI+ and NG2+/DAPI+ cells at 4 d after the C3a, C3a + C3aRa and C3aRa treatment when compared with the corresponding control (n = 5). For statistical analysis, one‐way ANOVA with Tukey's post hoc test was used for J–T and presented as the mean ± standard error of measurement (SEM). Scale bars: 20 μm. *P < 0.05, **P < 0.01.

Article Snippet: Image J software was used to quantify the intensity of the protein band (n = 6 each). table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Antibody Host Company Cat. No. RRID Application (concentration) C3a/C3 Chicken Abcam ab48581 AB_869211 WB (1:1000)/IF (1:200) C3aR Rabbit Abcam ab140777 AB_2687440 IF (1:100) C3aR Mouse Santa Cruz Biotechnology sc‐133172 AB_2066736 WB (1:1000)/IF (1:50) TMEM119 Rabbit Abcam ab209064 AB_2800343 IF (1:100) GFAP Mouse Abcam ab4648 AB_449329 IF (1:200) CNPase Mouse Cell Signaling Technology 5664 AB_10705455 WB (1:1000)/IF (1:200) MBP Rabbit Abcam ab62631 AB_956157 WB (1:1000)/IF (1:200) PLP Rabbit Abcam ab28486 AB_776593 WB (1:1000) MAG Mouse Abcam ab89780 AB_2042411 WB (1:1000) Olig1 Rabbit Abcam ab124908 AB_10972689 WB (1:1000) Olig2 Rabbit Abcam ab109186 AB_10861310 WB (1:1000)/IF (1:200) SOX10 Rabbit Abcam ab155279 AB_2650603 WB (1:1000) NG2 Mouse Abcam ab50009 AB_881569 WB (1:1000)/IF (1:200) CC1 Mouse Abcam ab16794 AB_443473 WB (1:1000)/IF (1:200) Ki‐67 Rabbit Cell Signaling Technology 9129 AB_2687446 IF (1:200) β‐catenin Rabbit Abcam ab32572 AB_725966 WB (1:1000) p‐β‐catenin Rabbit Cell Signaling Technology 9561 AB_331729 WB (1:1000) Tcf4 Mouse Novus H00006925‐M03 AB_2199267 WB (1:1000) Axin2 Rabbit Abcam ab32197 AB_2290204 WB (1:1000) β‐actin Mouse Cell Signaling Technology 58169 AB_2750839 WB (1:3000) GAPDH Rabbit Cell Signaling Technology 3683 AB_1642205 WB (1:3000) Open in a separate window Primary antibodies used in experiments.

Techniques: In Vitro, Immunofluorescence, Cell Culture, Expressing

Journal: Brain Pathology

Article Title: Complement C3a induces axonal hypomyelination in the periventricular white matter through activation of WNT/β‐catenin signal pathway in septic neonatal rats experimentally induced by lipopolysaccharide

doi: 10.1111/bpa.12798

Figure Lengend Snippet: C3a delays the differentiation and maturation of OPCs in vitro by activating the WNT/β‐catenin pathway. C3a administration activates the WNT/β‐catenin pathway in primary culture of OPCs. Panel A shows TCF4, β‐catenin, β‐catenin phosphorylation and AXIN2 immunoreactive bands after C3a treatment for 15 minutes, 30 minutes, 1, 2, and 4 h and the corresponding control (n = 6). Bar graphs (B–E) show the optical density of protein expression in A (n = 6). XAV939, an inhibitor of WNT/β‐catenin pathway, reverses the effect of C3a in primary culture of OPCs. Panel F shows the immunoreactive bands of PLP (26 kDa), MBP (33 kDa), CNPase (47 kDa), MAG (69 kDa), NG2 (251 kDa), CC1 (310 kDa), Olig1 (26 kDa), Olig2 (37 kDa), SOX10 (49 kDa) and β‐actin (42 kDa) after C3a, C3a + C3aRa, C3a + XAV939, XAV939 treatment and the corresponding control (n = 6). Bar graphs (G–O) show the optical density of protein expression in I (n = 6). For statistical analysis, one‐way ANOVA with Tukey's post hoc test was used for B–E and G–O and presented as the mean ± standard error of measurement (SEM). Scale bars: 20 μm. *P < 0.05, **P < 0.01.

Article Snippet: Image J software was used to quantify the intensity of the protein band (n = 6 each). table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Antibody Host Company Cat. No. RRID Application (concentration) C3a/C3 Chicken Abcam ab48581 AB_869211 WB (1:1000)/IF (1:200) C3aR Rabbit Abcam ab140777 AB_2687440 IF (1:100) C3aR Mouse Santa Cruz Biotechnology sc‐133172 AB_2066736 WB (1:1000)/IF (1:50) TMEM119 Rabbit Abcam ab209064 AB_2800343 IF (1:100) GFAP Mouse Abcam ab4648 AB_449329 IF (1:200) CNPase Mouse Cell Signaling Technology 5664 AB_10705455 WB (1:1000)/IF (1:200) MBP Rabbit Abcam ab62631 AB_956157 WB (1:1000)/IF (1:200) PLP Rabbit Abcam ab28486 AB_776593 WB (1:1000) MAG Mouse Abcam ab89780 AB_2042411 WB (1:1000) Olig1 Rabbit Abcam ab124908 AB_10972689 WB (1:1000) Olig2 Rabbit Abcam ab109186 AB_10861310 WB (1:1000)/IF (1:200) SOX10 Rabbit Abcam ab155279 AB_2650603 WB (1:1000) NG2 Mouse Abcam ab50009 AB_881569 WB (1:1000)/IF (1:200) CC1 Mouse Abcam ab16794 AB_443473 WB (1:1000)/IF (1:200) Ki‐67 Rabbit Cell Signaling Technology 9129 AB_2687446 IF (1:200) β‐catenin Rabbit Abcam ab32572 AB_725966 WB (1:1000) p‐β‐catenin Rabbit Cell Signaling Technology 9561 AB_331729 WB (1:1000) Tcf4 Mouse Novus H00006925‐M03 AB_2199267 WB (1:1000) Axin2 Rabbit Abcam ab32197 AB_2290204 WB (1:1000) β‐actin Mouse Cell Signaling Technology 58169 AB_2750839 WB (1:3000) GAPDH Rabbit Cell Signaling Technology 3683 AB_1642205 WB (1:3000) Open in a separate window Primary antibodies used in experiments.

Techniques: In Vitro, Expressing