Journal: Hearing research

Article Title: Culture media-based selection of endothelial cells, pericytes, and perivascular-resident macrophage-like melanocytes from the young mouse vestibular system

doi: 10.1016/j.heares.2016.12.012

Figure Lengend Snippet: Antibodies applied

Article Snippet: SOX10 , BiossInc, Woburn, MA , Cat# Bs-6449R , 1:50 , Rabbit polyclonal , Human, Mouse, Rat, Bovine.

Techniques:

Journal: BMC Neuroscience

Article Title: Therapeutic potential of hair follicle-derived stem cell intranasal transplantation in a rat model of ischemic stroke

doi: 10.1186/s12868-022-00732-w

Figure Lengend Snippet: Isolation and verification of HFSCs. In this study, the whisker pad ( A ) and hair follicles of the rat ( B ) were dissected. The hair bulges were isolated and explanted in collagen-coated plates ( C ). After 2–3 days of explantation, migrated cells were observed around the bulge ( C ). The immunostaining demonstrated the nestin ( D ) and SOX10 ( E ) expression that verify the migrated cells as HFSCs

Article Snippet: Identity of migrated HFSCs was assessed by immunostaining against nestin as a neural crest stem cell marker and SOX10 as a neural crest cells marker [ ] using mouse anti-nestin (1:50; Abcam, #ab6142) and rabbit anti-SOX10 (1:100; proteintech, 10422-1-AP) primary antibodies.

Techniques: Isolation, Whisker Assay, Immunostaining, Expressing

Journal: Scientific Reports

Article Title: Folate Metabolism Regulates Oligodendrocyte Survival and Differentiation by Modulating AMPKα Activity

doi: 10.1038/s41598-017-01732-1

Figure Lengend Snippet: DHFR is highly expressed in oligodendrocytes. ( A ) Spinal cords isolated from the wild type mice immunostained with DHFR antibody at P3, P8, P12 and P16. The ventral parts showed in the images. ( B , C ) The spinal cord of wild type mice at P15 is immunostained for DHFR and Sox10. The dorsal part showed in the image. A high magnification of ( B ) is shown in ( C ). Arrows indicate co-labeled cells. ( D , E ) The spinal cord of wild type mice at P15 is immunostained for DHFR, CC1, and PDGFRα, respectively. The ventral part showed in the image. A high magnification of ( E ) is shown in ( F ). Arrows indicated DHFR+/CC1+ cells. Arrowhead indicated DHFR+/PDGFRα+ cells. Scale bars: 100 μm ( A , B , D ), 40 μm ( C , E ).

Article Snippet: The primary antibodies were as follows: DHFR (Abcam ab85056, 1:500), Olig2 (Millipore Ab9610, 1:5000), CC1 (Calbiochem OP80, 1:50), MBP (Covance SMI-94R, 1:5000), PDGFRα (BD Bioscience 558774, 1:500), Sox10 (R&D systems NL2864R, 1:200), PLP (Millipore Mab388, 1:2000), NeuN (Millipore Mab377, 1:1000), cleaved-Caspase 3 (Cell signal 9661L, 1:1000), BrdU (BD pharmingen 555627, 1:200).

Techniques: Isolation, Labeling

Journal: Scientific Reports

Article Title: Folate Metabolism Regulates Oligodendrocyte Survival and Differentiation by Modulating AMPKα Activity

doi: 10.1038/s41598-017-01732-1

Figure Lengend Snippet: DHFR inhibition triggers evident oligodendrocyte damage and abnormal myelination. ( A , C ) qRT-PCR analysis of Dhfr mRNA level ( A ) or oligodendrocytes associated genes expression ( C ) in spinal cord from control and different doses of MTX-treated mice at P8. Data represents the mean ± S.D. (n > 3, * p < 0 . 05 compared with control, one-way ANOVA). ( B ) ELISA analysis of folate level in serum isolated from control and MTX-treated mice at P15. Data represents the mean ± S.D. (n = 5, * p < 0 . 05 compared with control, one-way ANOVA). ( D ) The spinal cords isolated from control and MTX-treated mice stained with DHFR and Sox10 antibodies at P8. The ventral parts showed in the images, and arrows indicate co-labeled cells. ( E ) Quantification of the numbers of DHFR+/Sox10+ cells per area (0.16 mm 2 ) in ( D ). Data represents the mean ± S.D. (n = 4, * p < 0 . 05 compared with control, student’s t test). ( F ) Immunohistochemistry analysis of spinal cords isolated from control and MTX-treated mice stained with MBP and PLP antibodies at P8. ( G ) Electron micrographs of spinal cord and optic nerve from control and MTX-treated mice at P15. Multilamellar myelin sheaths are compact around axons of control mice, whereas the myelin sheaths around axons from MTX-treated mice are essentially loose. Arrows represent the demyelinated axons, and asterisks indicate the unmyelinated axons. ( H ) G-ratio of individual axons as a function of axonal diameter in spinal cord and optic nerve from control and MTX-treated mice. (n > 100) Scale bars: 100 μm ( D ), 200 μm ( F ), 2 μm (( G ) left), 0.2 μm (( G ) right).

Article Snippet: The primary antibodies were as follows: DHFR (Abcam ab85056, 1:500), Olig2 (Millipore Ab9610, 1:5000), CC1 (Calbiochem OP80, 1:50), MBP (Covance SMI-94R, 1:5000), PDGFRα (BD Bioscience 558774, 1:500), Sox10 (R&D systems NL2864R, 1:200), PLP (Millipore Mab388, 1:2000), NeuN (Millipore Mab377, 1:1000), cleaved-Caspase 3 (Cell signal 9661L, 1:1000), BrdU (BD pharmingen 555627, 1:200).

Techniques: Inhibition, Quantitative RT-PCR, Expressing, Control, Enzyme-linked Immunosorbent Assay, Isolation, Staining, Labeling, Immunohistochemistry

Journal: Scientific Reports

Article Title: Folate Metabolism Regulates Oligodendrocyte Survival and Differentiation by Modulating AMPKα Activity

doi: 10.1038/s41598-017-01732-1

Figure Lengend Snippet: Oligodendrocyte defects induced by DHFR inhibition could be rescued by folate supplement. ( A ) Diagram shows the competition of folate and MTX on DHFR. Folate can block MTX entering cells by competitive combination of membrane receptor, leading to the reduction of DHFR inhibition. ( B ) qRT-PCR is carried out to analyze mRNA level of Dhfr , Mbp , Cnp , Myrf of spinal cords form control, MTX (4 mg/kg) and MTX (4 mg/kg) plus folate (10 mg/kg) mice at P8. Data represents the mean ± S.D. (n > 3, * p < 0 . 05 , one-way ANOVA). ( C ) Immunostaining using antibodies of Olig2, CC1 on spinal cord from control, MTX (4 mg/kg) and MTX (4 mg/kg) plus folate (10 mg/kg) mice at P8. The ventral parts showed in the images. ( D ) Quantification of the percentage of CC1+ cells among Olig2+ cells in ( C ). Data represents the mean ± S.D. (n = 5, * p < 0 . 05 , one-way ANOVA). ( E ) Immunohistochemistry using antibodies of Sox10 and PLP on spinal cord from control, MTX (4 mg/kg) and MTX (4 mg/kg) plus folate (10 mg/kg) mice at P8. ( F ) The number of quadrants with sparse staining for Sox10 and PLP in ( E ) scored and expressed as a percentage of the total number of quadrants. (n = 5, * p < 0 . 05 , one-way ANOVA). ( G , H ) Electron micrographs of spinal cord from control, MTX (4 mg/kg) and MTX (4 mg/kg) plus folate (10 mg/kg) mice at P15. ( H ) Shows the percentage of myelinated axons. Data represent the mean ± S.E.M from three animals (* p < 0 . 05 , one-way ANOVA). Scale bars: 50 μm ( C ), 200 μm ( E ), 2 μm ( G ).

Article Snippet: The primary antibodies were as follows: DHFR (Abcam ab85056, 1:500), Olig2 (Millipore Ab9610, 1:5000), CC1 (Calbiochem OP80, 1:50), MBP (Covance SMI-94R, 1:5000), PDGFRα (BD Bioscience 558774, 1:500), Sox10 (R&D systems NL2864R, 1:200), PLP (Millipore Mab388, 1:2000), NeuN (Millipore Mab377, 1:1000), cleaved-Caspase 3 (Cell signal 9661L, 1:1000), BrdU (BD pharmingen 555627, 1:200).

Techniques: Inhibition, Blocking Assay, Membrane, Quantitative RT-PCR, Control, Immunostaining, Immunohistochemistry, Staining