Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Blood–spinal cord barrier pericyte reductions contribute to increased capillary permeability

doi: 10.1038/jcbfm.2012.113

Figure Lengend Snippet: Pericyte number and coverage is reduced along blood–spinal cord barrier. ( A ) Confocal microscopy analysis of CD13-positive pericytes (green) and collagen IV-positive capillary profiles (red) in 2-month-old wild-type mouse cortex, caudate, and hippocampal brain regions and cervical, thoracic, and lumbar spinal cord regions (anterior horns). ( B ) Quantification of regional CD13-positive pericyte coverage of collagen IV-positive brain and spinal cord capillaries. Mean±s.e.m., n =5 to 7 animals per group. # P <0.05 when compared with the brain regions; * P <0.05. ( C ) Quantification of regional CD13-positive pericyte cell number normalized to collagen IV-positive capillary surface area in the brain and spinal cord. Mean±s.e.m., n =5 to 7 animals per group. # P <0.05 when compared with the brain regions; * P <0.05. ( D ) Confocal microscopy analysis of platelet-derived growth factor receptor β (PDGFR β )-positive pericytes (green) and lectin capillary profiles (red) in 2-month-old wild-type mouse cortex, caudate, and hippocampal brain regions and cervical, thoracic, and lumbar spinal cord regions. ( E ) Quantification of regional PDGFR β -positive pericyte coverage of lectin-positive brain and spinal cord capillaries. Mean±s.e.m., n =7 animals per group. # P <0.05 when compared with the brain regions; * P <0.05. ( F ) Quantification of regional PDGFR β -positive pericyte cell number normalized to lectin-positive capillary surface area in the brain and spinal cord. Mean±s.e.m., n =7 animals per group. # P <0.05 when compared with the brain regions; * P <0.05.

Article Snippet: Sections were then incubated in the following primary antibodies: goat α -mouse CD13 (10 μ g/mL, R&D Systems), mouse α -mouse desmin (1:100, Dako, Carpinteria, CA, USA), rabbit α -mouse collagen IV (1:200, EMD Millipore), rabbit α -human fibrin which cross reacts with mouse fibrin (1:500, Dako), goat α -mouse thrombin (1:50, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse α -mouse zonula occludens-1 (ZO-1) (1:50, Invitrogen), mouse α -mouse occludin (1:50, Invitrogen), mouse α -mouse neuronal-specific antigen A60 (NeuN) (1:100, EMD Millipore) and goat anti-mouse ChAT (choline acetyl transferase) (1:50, EMD Millipore).

Techniques: Confocal Microscopy, Derivative Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Blood–spinal cord barrier pericyte reductions contribute to increased capillary permeability

doi: 10.1038/jcbfm.2012.113

Figure Lengend Snippet: Subregional variation in spinal cord pericyte number and coverage between anterior horn gray matter and lateral and dorsal funiculi. ( A ) Confocal microscopy analysis of CD13-positive pericytes (green) and collagen IV-positive capillary profiles (red) in 2-month-old wild-type mouse anterior horn gray matter or dorsal columns for cervical thoracic and lumbar spinal cord. ( B ) Quantification of CD13-positive pericyte coverage of collagen IV-positive capillaries in anterior horn gray matter (gray) or dorsal and lateral funiculi (white) from cervical, thoracic, and lumbar spinal cord. Mean±s.e.m., n =5 to 7 animals per group; * P <0.05. ( C ) Quantification of CD13-positive pericyte cell number normalized to lectin-positive capillary surface area in anterior horn gray matter (gray) or dorsal and lateral funiculi (white) from cervical, thoracic, and lumbar spinal cord. Mean±s.e.m., n =5 to 7 animals per group; * P <0.05. ( D ) Quantification of platelet-derived growth factor receptor β (PDGFR β )-positive pericyte coverage of lectin-positive capillaries in anterior horn gray matter (gray) or dorsal and lateral funiculi (white) from cervical, thoracic, and lumbar spinal cord. Mean±s.e.m., n =7 animals per group; * P <0.05. ( E ) Quantification of PDGFR β -positive pericyte cell number normalized to lectin-positive capillary surface area in anterior horn gray matter (gray) or dorsal and lateral funiculi (white) from cervical, thoracic, and lumbar spinal cord. Mean±s.e.m., n =7 animals per group; * P <0.05.

Article Snippet: Sections were then incubated in the following primary antibodies: goat α -mouse CD13 (10 μ g/mL, R&D Systems), mouse α -mouse desmin (1:100, Dako, Carpinteria, CA, USA), rabbit α -mouse collagen IV (1:200, EMD Millipore), rabbit α -human fibrin which cross reacts with mouse fibrin (1:500, Dako), goat α -mouse thrombin (1:50, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse α -mouse zonula occludens-1 (ZO-1) (1:50, Invitrogen), mouse α -mouse occludin (1:50, Invitrogen), mouse α -mouse neuronal-specific antigen A60 (NeuN) (1:100, EMD Millipore) and goat anti-mouse ChAT (choline acetyl transferase) (1:50, EMD Millipore).

Techniques: Confocal Microscopy, Derivative Assay

Journal: Journal of Cerebral Blood Flow & Metabolism

Article Title: Blood–spinal cord barrier pericyte reductions contribute to increased capillary permeability

doi: 10.1038/jcbfm.2012.113

Figure Lengend Snippet: Exacerbation of pericyte deficiency leads to overt blood–spinal cord barrier disruption and leakage of endogenous plasma proteins. ( A ) Confocal microscopy analysis of CD13-positive pericytes (green) and collagen IV-positive capillary profiles (red) in 6-month-old Pdgfrβ +/+ and Pdgfrβ F7/F7 mouse cervical, thoracic, and lumbar spinal cord anterior horn. ( B ) Quantification of regional CD13-positive pericyte coverage of collagen IV-positive anterior horn spinal cord capillaries. Mean±s.e.m., n =3 animals per group; * P <0.05. ( C ) Representative confocal microscopy analysis of IgG (green), fibrin (red), and lectin-positive capillaries (blue) in 6-month-old Pdgfrβ +/+ mouse lumbar anterior horn and anterior horn from Pdgfrβ F7/F7 cervical, thoracic, and lumbar spinal cord. ( D , E ) Quantification of IgG ( D ) and fibrin ( E ) extravascular deposits in the spinal cord regions in tissue sections from 2-month-old B6SJL/F1 Pdgfrβ +/+ , 6-month-old Pdgfrβ +/+ 129S1/SvlmJ, and 6-month-old Pdgfrβ F7/F7 mice. Mean±s.e.m., n =3 animals per group; * P <0.05. ( F , G ) Confocal microscopy analysis of thrombin (white) ( F ) or fibrin (red) ( G ) and SMI-311-positive neurons (blue) in 6-month-old Pdgfrβ +/+ and Pdgfrβ F7/F7 lumbar spinal cord. Arrows indicate neuronal accumulation.

Article Snippet: Sections were then incubated in the following primary antibodies: goat α -mouse CD13 (10 μ g/mL, R&D Systems), mouse α -mouse desmin (1:100, Dako, Carpinteria, CA, USA), rabbit α -mouse collagen IV (1:200, EMD Millipore), rabbit α -human fibrin which cross reacts with mouse fibrin (1:500, Dako), goat α -mouse thrombin (1:50, Santa Cruz Biotechnology, Santa Cruz, CA, USA), mouse α -mouse zonula occludens-1 (ZO-1) (1:50, Invitrogen), mouse α -mouse occludin (1:50, Invitrogen), mouse α -mouse neuronal-specific antigen A60 (NeuN) (1:100, EMD Millipore) and goat anti-mouse ChAT (choline acetyl transferase) (1:50, EMD Millipore).

Techniques: Disruption, Clinical Proteomics, Confocal Microscopy

Journal: Scientific Reports

Article Title: SPOCK1 is a novel inducer of epithelial to mesenchymal transition in drug-induced gingival overgrowth

doi: 10.1038/s41598-020-66660-z

Figure Lengend Snippet: SPOCK1 overexpression induces EMT-like changes at histological and molecular levels in mice gingiva. (a) Relative protein expression of E-cadherin and vimentin was assessed by western blotting in TG and WT mice gingival tissues with GAPDH as an internal control. Data shown are presented as means ± SEM; *P < 0.05 and ***P < 0.001 compared with WT controls analyzed by the Mann-Whitney U test, (n = 5 per group). ( b) IF staining of E-cadherin, vimentin and α E-catenin of TG and WT specimens at x40 magnification. The cells in the epithelium express less E-cadherin and α E-catenin, while cells within the connective and the basal layer of the epithelium expressed more vimentin in TG mouse gingiva compared to WT gingiva. scale bar = 100 μm. ( c) Relative protein expression levels of MMP-9 and MMP-2 were measured by western blotting in TG and WT mouse gingiva, with GAPDH used as an internal control. Data shown are presented as means ± SEM; ***P < 0.001 compared with WT controls analyzed by the Mann-Whitney U test, (WT n = 6, TG n = 9). ( d) Relative protein expression of TGF-β1, CTGF, and SLUG measured by western blotting in TG and WT mouse gingiva. GAPDH was used as internal control. Data shown are presented as means ± SEM; ***P < 0.001 compared with WT controls analyzed by the Mann-Whitney U test.

Article Snippet: Sections were then incubated with SPOCK1 mouse anti-human (1:200; R&D SYSTEMS), TGF-β1 rabbit anti-human (1:200; Abcam) Collagen IV rabbit anti-mouse (2:200; Abcam, #ab6568), E-cadherin rabbit anti-mouse (1:200; Cell Signaling Technology, Beverly, MA, #24E10), vimentin mouse anti-mouse (4:200; Santa Cruz), andαE-catenin rabbit anti-mouse (6:200; Proteintech, #12831-1-AP) primary antibodies at 4 °C overnight, the sections washed and incubated with Alexa 488-conjugated goat anti-rabbit IgG (1:200; Abcam, #ab150077) or Alexa 647-conjugated goat anti-mouse IgG (1:200; Abcam, #ab150115).

Techniques: Over Expression, Expressing, Western Blot, Control, MANN-WHITNEY, Staining