Journal: Brain research

Article Title: Endothelins as pronociceptive mediators of the rat trigeminal system: role of ETA and ETB receptors.

doi: 10.1016/j.brainres.2010.04.075

Figure Lengend Snippet: Fig. 2 – Expression of immunoreactive ETA and ETB receptors in fluorogold-labeled trigeminal ganglion neurons. Animals received fluorogold (FG, 4% solution) injected s.c. into the upper lip. After 5–7 days, FG-injected rats were sacrificed and both TG removed and processed for ETA and ETB receptor immunohistochemistry. In panel A, images on the right depict FG-labeled TG neurons; middle images show ETA and ETB receptor staining and images on the left show expression of ETA and ETB receptors on some FG positive neurons (arrows on merged images). The asterisk indicates ETB receptor expression on satellite glial cells. Tissue sections incubated in the absence of primary or secondary antibodies did not display any fluorescence (not shown). Images are representative of at least 3 sections from each ganglion, taken from 3 to 4 distinct animals. Panel B illustrates the size–frequency histogram of ETA and ETB positive neurons within the entire trigeminal ganglion (n=1460 neurons).

Article Snippet: For double-labeling experiments, slides were incubated simultaneously with the anti-ETA or anti-ETB receptor antibody and goat anti-TRPV1 receptor antibody (1:200; Santa Cruz Biotechnology, CA, USA).

Techniques: Expressing, Labeling, Injection, Immunohistochemistry, Staining, Incubation, Fluorescence

Journal: Brain research

Article Title: Endothelins as pronociceptive mediators of the rat trigeminal system: role of ETA and ETB receptors.

doi: 10.1016/j.brainres.2010.04.075

Figure Lengend Snippet: Fig. 3 – Co-localization of ETA and ETB receptors with TRPV1. Sections were labeled for TRPV1 receptors (in red, left images) and ETA or ETB receptors (ETAR and ETBR, in green). Images on the right depict co-localization of TRPV1 receptors with ETAR and ETBR (arrows on merged images). Tissue sections incubated in the absence of primary or secondary antibodies did not display any fluorescence (not shown). Images are representative of at least 3 sections from each ganglion taken from 3 to 4 distinct animals. Panel B illustrates the percentage of neurons immunoreactive for TRPV1, ETA (ETAR) and ETB receptors (ETBR), as well as the percentage of TRPV1 positive neurons that also express ETAR and ETBR (n=1102 neurons).

Article Snippet: For double-labeling experiments, slides were incubated simultaneously with the anti-ETA or anti-ETB receptor antibody and goat anti-TRPV1 receptor antibody (1:200; Santa Cruz Biotechnology, CA, USA).

Techniques: Labeling, Incubation, Fluorescence

Journal: Brain research

Article Title: Endothelins as pronociceptive mediators of the rat trigeminal system: role of ETA and ETB receptors.

doi: 10.1016/j.brainres.2010.04.075

Figure Lengend Snippet: Fig. 7 – Influence of ETA and ETB receptor antagonists on ET-1-induced nociceptive responses. BQ-123 or BQ-788 (selective ETA and ETB receptor antagonists, each at 10 nmol/site) were administered 30 min before ipsilateral ET-1 (10 pmol/site) injection into the upper lip (A) or TMJ (B). Each value represents the mean±s.e.m. of the total facial grooming time (A) or the sum of nociceptive behaviors (B) recorded from 6 to 8 animals. Asterisks denote P≤0.05 relative to the PBS plus ET-1-injected group (ANOVA followed by Bonferroni's test).

Article Snippet: For double-labeling experiments, slides were incubated simultaneously with the anti-ETA or anti-ETB receptor antibody and goat anti-TRPV1 receptor antibody (1:200; Santa Cruz Biotechnology, CA, USA).

Techniques: Injection

Journal: PLoS Neglected Tropical Diseases

Article Title: Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment

doi: 10.1371/journal.pntd.0009362

Figure Lengend Snippet: MPAs were infected with ZIKV at MOI of 2 for 0, 24, and 48 h, respectively. (A) ZIKV NS5 RNA was determined by qPCR analysis in MPAs. (B) The mRNA expressions of PTBP1 , GHR , LIF , PTBP3 , EDNRB , and MBP were determined by qPCR. The data represent the relative expression of target genes normalized to GAPDH as a reference internal control. ( C ) The protein expressions of PTBP1, LIF, GHR, EDNRB, and MBP were determined by Western blot analyses. The β-actin was selected as an internal control. (D and E) Confocal microscopy images from MPAs stained with PTBP1, GHR, LIF, EDNRB, and MBP (green) for target genes and DAPI (blue) for nuclei, scale bar = 20 μm ( D ). Relative protein expression to control is quantified using Image J software (E) . Data are shown as fold change from mock control. All the experiments were performed in triplicate, non-significant (ns); P < 0.05 (*); P < 0.01 (**); P < 0.001 (***). Student’s t -test. The underlying numerical data for “Fig 4A, 4B, and 4E” is provided in the supporting file “ ”.

Article Snippet: Rabbit anti- EDNRB (A2980), MBP (A11162), LIF (A1288), GHR (A14735) and β-actin antibodies were purchased from ABclonal Technology (Wuhan, China).

Techniques: Infection, Expressing, Western Blot, Confocal Microscopy, Staining, Software

Journal: PLoS Neglected Tropical Diseases

Article Title: Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment

doi: 10.1371/journal.pntd.0009362

Figure Lengend Snippet: MPAs were infected with ZIKV at MOI of 2 for 0 and 24 h, respectively. ( A - F ) The expression patterns of top 6 shortlisted genes: PTBP1 (A), GHR (B), LIF (C), PTBP3 (D), EDNRB (E), and MBP (F), were measured by RNA-seq and qRT-PCR (qPCR), respectively. GAPDH was used as an internal control in qRT-PCR. The individual bars represent the qPCR data as the means ± SD, n = 3. The underlying numerical data for “Fig 5A–5F” is provided in the supporting file “ ”.

Article Snippet: Rabbit anti- EDNRB (A2980), MBP (A11162), LIF (A1288), GHR (A14735) and β-actin antibodies were purchased from ABclonal Technology (Wuhan, China).

Techniques: Infection, Expressing, RNA Sequencing Assay, Quantitative RT-PCR

Journal: PLoS Neglected Tropical Diseases

Article Title: Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment

doi: 10.1371/journal.pntd.0009362

Figure Lengend Snippet: U251 cells were infected with ZIKV at MOI of 2 for indicated periods. (A) Cell viability was measured by the CCK-8 assay. Data represent as fold change compared to mock-treated cells. (B) ZIKV NS5 RNA was determined by qPCR with GAPDH as an internal control, and (C) viral dsRNA was immunoprobed with dsRNA-antibody (red), along with GFAP (green) for astrocytes and DAPI (blue) for nuclei by Immunofluorescent (IF), scale bar = 20 μm. (D and E) The mRNA expressions of PTBP1, GHR, LIF, PTBP3, EDNRB, and MBP were determined by qPCR ( D ) and protein expressions of PTBP1, GHR, LIF, EDNRB, and MBP was determined by Western blot analyses ( E ), respectively. In the qPCR analysis, the data represent the relative expression of target genes normalized to GAPDH as a reference internal control. In Western blot analysis, the β-actin was selected as an internal control. (F and G) Confocal microscopy images from MPAs stained with PTBP1, GHR, LIF, EDNRB, and MBP (green) for target genes and DAPI (blue) for nuclei, scale bar = 20 μm ( F ). Relative protein expression to control is quantified using Image J software (G) . Data are shown as fold change from mock control. All the experiments were performed in triplicate, non-significant (ns); P < 0.05 (*); P < 0.01 (**); P < 0.001 (***). Student’s t -test. The underlying numerical data for “Fig 6A, 6B, 6D, and 6G” is provided in supporting file “ ”.

Article Snippet: Rabbit anti- EDNRB (A2980), MBP (A11162), LIF (A1288), GHR (A14735) and β-actin antibodies were purchased from ABclonal Technology (Wuhan, China).

Techniques: Infection, CCK-8 Assay, Western Blot, Expressing, Confocal Microscopy, Staining, Software

Journal: PLoS Neglected Tropical Diseases

Article Title: Zika virus dysregulates the expression of astrocytic genes involved in neurodevelopment

doi: 10.1371/journal.pntd.0009362

Figure Lengend Snippet: The neurotropic ZIKV favorably infects and replicates in astrocytes of the fetal brain cerebral cortex. ZIKV can alter the astrocytic genes including PTBP1, LIF, PTBP3, GHR, EDNRB, and MBP, which are involved in various functions leading to neurological conditions such as microcephaly and brain development impairment. The numerical data used in all figures are included in .

Article Snippet: Rabbit anti- EDNRB (A2980), MBP (A11162), LIF (A1288), GHR (A14735) and β-actin antibodies were purchased from ABclonal Technology (Wuhan, China).

Techniques:

Journal: BMC Neuroscience

Article Title: Upregulation of the endothelin A (ET A ) receptor and its association with neurodegeneration in a rodent model of glaucoma

doi: 10.1186/s12868-017-0346-3

Figure Lengend Snippet: Overexpression of the ET A receptor can increase in ET B receptor expression. a Representative images. Immunoblot showing the expression of ET A and ET B receptors in the plasma membrane fraction of 661W stable clones overexpressing the ET A receptor. b Immunoblot densitometry showing fold change in protein expression of both ET A and ET B receptors in 661W cells overexpressing the ET A receptor compared to empty vector cells. Protein normalized to calnexin expression. Bars represent mean ± SEM (n = 5). Asterisks indicated statistical significance *p < 0.05 by student’s t-test

Article Snippet: Primary antibodies used to probe blots were rabbit anti-ET A (1:1000; Sigma), rabbit anti-ET B (1:10,000, Antibody Research Corporation), rabbit anti-Calnexin (1:1000, Cell Signaling) and mouse calnexin (1:1000, Cell Signaling).

Techniques: Over Expression, Expressing, Western Blot, Clone Assay, Plasmid Preparation

Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: ERK1/2 inhibition attenuates cerebral blood flow reduction and abolishes ET(B) and 5-HT(1B) receptor upregulation after subarachnoid hemorrhage in rat.

doi: 10.1038/sj.jcbfm.9600236

Figure Lengend Snippet: Figure 4 Effect of treatment with ERK1/2 inhibitor SB386023-b in cerebral arteries on the mRNA levels of 5-HT1B and ETB receptors after experimental induced SAH in rats. (A) ETB and (B) 5-HT1B. There are upregulations of the ETB and 5-HT1B receptor mRNA levels in the SAH compared with the sham- operated rats. Treatment with SB386023-b prevented the upregulations. Data were obtained by real-time PCR and are expressed as mean7s.e.m. values relative to EF-1 mRNA levels, n = 7 to 10, *Pr0.05.

Article Snippet: The first antibodies used were rabbit antihuman ETB (IBL, Stockholm, Sweden, 16207), diluted 1:400, goat anti-mouse 5-HT1B (Santa Cruz Biotechnologies, Santa Cruz, CA, USA, sc-1461), diluted 1:100, mouse anti-rat CD31 (Serotec, Hamar, Norway, MCA1746), diluted 1:200, and mouse anti-rat smooth muscle actin (Serotec, MCA1905T), diluted 1:100.

Techniques: Real-time Polymerase Chain Reaction

Journal: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism

Article Title: ERK1/2 inhibition attenuates cerebral blood flow reduction and abolishes ET(B) and 5-HT(1B) receptor upregulation after subarachnoid hemorrhage in rat.

doi: 10.1038/sj.jcbfm.9600236

Figure Lengend Snippet: Figure 5 Sections from the BA showing ETB and 5-HT1B immunoreactivity in the smooth muscle cell layer. (A) ETB; sham, (B) ETB; SAH, (C) ETB; SAH treated with SB386023-b, (D) 5-HT1B; sham, (E) 5-HT1B; SAH, (F) 5-HT1B; SAH treated with SB386023-b. There are increased expressions of the ETB and 5-HT1B receptor protein levels in the SAH compared with the sham operated rats. Treatment with SB386023-b prevented the increased protein expression in the smooth muscle cells. Data were obtained with confocal microscopy.

Article Snippet: The first antibodies used were rabbit antihuman ETB (IBL, Stockholm, Sweden, 16207), diluted 1:400, goat anti-mouse 5-HT1B (Santa Cruz Biotechnologies, Santa Cruz, CA, USA, sc-1461), diluted 1:100, mouse anti-rat CD31 (Serotec, Hamar, Norway, MCA1746), diluted 1:200, and mouse anti-rat smooth muscle actin (Serotec, MCA1905T), diluted 1:100.

Techniques: Expressing, Confocal Microscopy