Journal: Neural Regeneration Research

Article Title: Inhibition of LncRNA Vof-16 expression promotes nerve regeneration and functional recovery after spinal cord injury

doi: 10.4103/1673-5374.314322

Figure Lengend Snippet: Transplantation of lncRNA Vof-16 knockdown lentivirus promotes the outgrowth of regenerative fibers in rats with SCI. After SCI, rats received 10 µL PBS or lncRNA Vof-16 lentivirus in the lesion gap. (A) lncRNA Vof-16 was upregulated after SCI. (A) Representative immunofluorescent double staining with NF-200 (regenerative nerve fiber marker, green) and CD31 (vascular marker, red) in the lesion of rats from different treatment groups 7 days post-injury. Knockdown of lncRNA Vof-16 by lentivirus promoted the regeneration of nerve fibers after SCI. The white dotted line represents the spinal cord injury cavity region. The red box represents the local higher magnification view. The white arrows indicate blood vessels (CD31 + ). Scale bars: 100 µm. (B–D) The quantity (B) and relative area (C) of nerve regeneration fibers and the relative area of angiogenesis (D) were counted and plotted. Data are shown as the mean ± SD ( n = 4/group). The experiment was repeated three times. ** P < 0.01, *** P < 0.001, vs . PBS group (one-way analysis of variance, followed by Tukey’s post hoc test). Sham: sham operation group; PBS: PBS group; NC-Knockdown: knockdown lentiviral vector negative control group; Knockdown: lncRNA Vof-16 knockdown lentivirus group; NC-Overexpression: overexpression lentivirus vector negative control group; Overexpression: lncRNA Vof-16 overexpression lentivirus group. lncRNA: long non-coding RNA; NF-200: neurofilament protein-200; SCI: spinal cord injury.

Article Snippet: The tissue sections were then incubated with primary antibodies specific to neurofilament protein-200 (NF-200; Cat# N4142; monoclonal antibody; rabbit anti-rat; 1:400; Sigma), CD31 (Cat# AF3628; polyclonal antibody; goat anti-rat; 1:200; R&D Systems, Minneapolis, MN, USA), or NeuN (monoclonal antibody; mouse anti-rat; 1:200; Millipore) overnight at 4°C.

Techniques: Transplantation Assay, Knockdown, Double Staining, Marker, Plasmid Preparation, Negative Control, Over Expression

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Localization of CD31 (+) and endomucin (+) vessels. ( A ) Stage of tooth crown formation at 1 week. Endomucin (+) vessels (red) mainly exist in the dental pulp. ( B ) Higher magnifications of the boxed regions in A . Both CD31(+) and endomucin (+) vessels (yellow) are narrow and located beneath the coronal dentin. ( C ) Higher magnifications of the boxed regions in B . Merged immunoreaction (yellow) is localized at thin vessels in the odontoblast layer. ( D ) Dental pulp at 4 weeks. Both CD31 (+) and endomucin (+) vessels decreased in the crown and increased in the root of dental pulp at 4 weeks. ( E ) Higher magnifications of the boxed regions in D . Thin vessels are colored in yellow, and express both CD31 (+) and endomucin (+) and located near the radicular dentin (arrowheads). ( F ) Higher magnifications of the boxed regions in E . Note the merged immunoreaction (yellow) shows mesh-like pattern. ( G ) Dental pulp at 12 weeks. Root canals at 12 week are narrower than those at 4 week. ( H ) Higher magnifications of the boxed regions in G . CD31 (+)-immunoreaction is mainly seen, and Both CD31 (+) and endomucin (+) vessels are faintly seen in dental pulp (arrowhead). ( I ) Higher magnifications of the boxed regions in H . CD31 (+) vessels are seen beneath the odontoblast layer. ( J ) Dental pulp at 56 weeks. Few CD31 (+) and endomucin (+) vessels were detected in dental pulp. ( K ) Higher magnifications of the boxed regions in J . Main vessels are positive for CD31 in the coronal pulp. ( L ) Higher magnifications of the boxed regions in K . Immuno-reaction for endomucin was separately seen at the CD31(+) vessels. Green: CD31 (+), Red: Endomucin (+), Yellow: CD31 (+) and endomucin (+). Arrowheads: CD31 (+) and endomucin (+) vessels. Den: dentin; Ob: odontoblast layer. Bars = 200 μm ( A , D , G , J ), 50 μm ( B , E , H , K ) and 15 μm ( C , F , I , L ).

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques:

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Ratio of CD31 (+) and endomucin (+) vessels. Data represent means ± standard deviations (SD). Four mice were used for statistical analysis in each group. These ratios 12 and 56 weeks were significantly lower than those 1 and 4 weeks. * Statistical significance ( P < 0.05)

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques:

Journal: Acta Histochemica et Cytochemica

Article Title: Localization of Both CD31- and Endomucin-Expressing Vessels in Mouse Dental Pulp

doi: 10.1267/ahc.24-00009

Figure Lengend Snippet: Calcein labeling and endomucin (+) vessels. ( A ) At 4 weeks after birth. Dentin with a large distance between the two calcein-lines was observed in the crown and root dentin. ( B ) Higher magnification of the boxed regions in A . Both CD31(+) and endomucin (+) vessels colored in white were thin and present directly beneath calcein-labeled dentin at all sites. ( C ) At 8 weeks after birth. Although the lines were present in root dentin, they were negligible in coronal dentin. The distance between the two lines at 8 weeks were narrower than that at 4 weeks. Both CD31 and endomucin (+) vessels were scattered in coronal dental pulp. ( D ) Higher magnification of the boxed regions in C . Both CD31 and endomucin (+) vessels were located at the odontoblast layer in root pulp. ( E ) At 12 weeks after birth. Calcein-labeled areas were only present in newly formed apical cementum, and were faint or not seen at coronal and root dentin. ( F ) Higher magnification of the boxed regions in E . CD31(+) and ndomucin (+) vessels were seen neat the newly formed cementum and faintly detected in dental pulp. Den: dentin, Cem: cementum, Ob: odontoblast layer, Pu: dental pulp. Bars = 200 μm ( A , C , E ), 30 μm ( B , D , F ). Green: calcein labeling, Cyan: CD31(+) vessel, Red: endomucin (+) vessel, White: both CD31(+) and endomucin(+) vessel, Blue: nuclear.

Article Snippet: Sections of decalcified samples were incubated with an Alexa Fluor 488-conjugated anti-CD31/PECAM-1 goat polyclonal antibody (dilution 1:100, FAB3628G-10, R&D Systems, Minneapolis, MN, USA) and a rat monoclonal antibody against mouse endomucin (dilution 1:200, sc-65495, Santa Cruz Biotechnology Inc., Dallas, TX, USA) at 20 ± 5°C for 8 hr or 4°C overnight in the dark.

Techniques: Labeling

Journal: Nature Metabolism

Article Title: SIRT2 regulates extracellular vesicle-mediated liver–bone communication

doi: 10.1038/s42255-023-00803-0

Figure Lengend Snippet: a , The blood concentration of sEVs at different time points after tail vein injection of sEVs. n = 3 mice. One technical replicate of 3 biological replicates. b , Immunofluorescence analysis of the distribution of sEV-LRG1-GFP (green) in RANKL-induced BMDMs at 12 h and 24 h after supplementing sEVs labeled with PKH26 (red) in supernatant (scale bar, 20 µm). c , Enrichment of signaling pathway of sEV-LRG1 binding proteins in DAVID Bioinformatics database. d , IHC detection of CD31 in the paraffin-embedded bone section of distal femur of aged LoxP and SIRT2 -KO hep mice (scale bar, 100μm). e , Quantification of CD31 positive vessels area (aged LoxP mice: n = 10 and aged SIRT2 -KO hep mice: n = 12). One technical replicate of 10 (LoxP mice) or 12 ( SIRT2 -KO hep mice) biological replicates for each group. f , Western blot analysis of p65 protein levels in the RAW 264.7 cells overexpressed p65. n = 3 biologically independent experiments. g , HEK293T cells transfected with LPHN2 plasmid were treated with LRG1-sEVs (4 μg/ml) and then immunofluorescence colocalization analysis of sEV-LRG1 and LPNH2 was shown (scale bar, 20 µm). n = 2 biologically independent experiments. Data are presented as mean ± SD. with biologically individual data points shown. P values are determined by one-way ANOVA followed by Tukey’s test ( a ) and unpaired two-tailed Student’s t -test ( e ). n.s., not significant.

Article Snippet: The corresponding antibodies (CD31 goat pAb, 1:200 dilution, ServiceBio, GB13063) and HRP-conjugated rabbit anti-goat IgG (H+L) (1:200 dilution, ServiceBio, GB23204).

Techniques: Concentration Assay, Injection, Immunofluorescence, Labeling, Binding Assay, Western Blot, Transfection, Plasmid Preparation, Two Tailed Test