Journal: Acta biochimica Polonica

Article Title: Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study.

doi: 10.18388/abp.2017_1520

Figure Lengend Snippet: Figure 3. Survival curves for Colo-205 exposed to FAAH inhibi tor PF-3845 alone or co-incubated with selected antagonists: AM 251 (CB1 receptor antagonist), AM 630 (CB2), SB 366791 (TRPV1), RN 1734 (TRPV4) and G-15 (GPR30). Each point represents mean of 5 independent determinations. Standard deviations were excluded for clarity and did not exceed 12% of the mean value for each point.

Article Snippet: The membrane was then washed in TBS-T and hybridized with primary rabbit CB1 and CB2 polyclonal antibodies (Santa Cruz Biotechnology, Dallas, TX, USA), which were diluted to a suitable concentration (1:200 – 1:500) in TBS-T, for 16 h. Additionally, the membranes were incubated with a goat polyclonal antibody specific for β-actin (1:100 000 dilution) to use as a loading control.

Techniques: Incubation

Journal: Acta biochimica Polonica

Article Title: Fatty acid amide hydrolase (FAAH) inhibitor PF-3845 reduces viability, migration and invasiveness of human colon adenocarcinoma Colo-205 cell line: an in vitro study.

doi: 10.18388/abp.2017_1520

Figure Lengend Snippet: Figure 4. Effect of PF-3845 on CB1 and CB2 receptor expression in the Colo-205 cells. (A) Cells were treated with PF-3845 for 48 h and subjected to Western blot analysis. PF-3845 decreased expression of CB1 and in creased expression of CB2 receptor. (B) β-actin in the cell lysate is shown as control. Each blot is representative of at least three oth er blots. Density values of the bands are graphically expressed rel ative to control. Data are shown as mean ± S.D. (n=4). Significance of differences between means: *p<0.05 versus control; ***p<0.001 versus control. Figure 5. Inhibition of Colo-205 cell migration by PF-3845 in the concentration range of 0 to 100 µM. Migration studies were conducted using BD FalconTM 24-Multiwell Insert plate system. Total area of inserts with crystal violet-stained cells was photographed and counted using the NIH ImageJ analy sis software. (A) Representative images obtained for Colo-205 are shown above graphs. Initial magnification: x 200. (B) Each value on the graph represents the mean value ± S.D., n=3 independent ex periments. Significance of differences between means: ***p<0.001 versus control.

Article Snippet: The membrane was then washed in TBS-T and hybridized with primary rabbit CB1 and CB2 polyclonal antibodies (Santa Cruz Biotechnology, Dallas, TX, USA), which were diluted to a suitable concentration (1:200 – 1:500) in TBS-T, for 16 h. Additionally, the membranes were incubated with a goat polyclonal antibody specific for β-actin (1:100 000 dilution) to use as a loading control.

Techniques: Expressing, Western Blot, Control, Inhibition, Migration, Concentration Assay, Staining, Software

Journal: Arthritis Research & Therapy

Article Title: Elevated serum immunoglobulin level predicts high risk of 1-year recurrence in patients with Takayasu arteritis

doi: 10.1186/s13075-023-03016-8

Figure Lengend Snippet: CD138 expressions in the aortic wall of TAK and control group. A CD138 staining in the aortic wall of TAK patients and control group (scale bars, 100 μm; n = 3 in control group, n = 4 in TAK group). B The quantification of CD138 staining in the two groups ( n = 3 in control group, n = 4 in TAK group)

Article Snippet: Then, they were stained with primary antibody for CD138 (Item No.ab46506, Abcam, Cambridge, CB2 0AX, UK) overnight at 4 °C and secondary antibody for 0.5–1 h at room temperature and detected with 3,3′-diaminobenzidine.

Techniques: Control, Staining

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Spatio-temporal expression patterns of anandamide-binding receptors in rat implantation sites: evidence for a role of the endocannabinoid system during the period of placental development

doi: 10.1186/1477-7827-7-121

Figure Lengend Snippet: Primer sequences, gene accession number and Q-PCR conditions.

Article Snippet: The separated proteins were transferred onto a nitrocellulose membrane in 25 mM Tris-HCl, 250 mM glycine, 20% methanol (v/v), pH 8.3, for 2 h at 200 V. The membranes were incubated with a 1:100 dilution of polyclonal antibodies against CB1 (sc-20754), CB2 (sc-25494) or TRPV1 (sc-12498) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) in blocking solution overnight at 4°C.

Techniques:

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Spatio-temporal expression patterns of anandamide-binding receptors in rat implantation sites: evidence for a role of the endocannabinoid system during the period of placental development

doi: 10.1186/1477-7827-7-121

Figure Lengend Snippet: Anandamide binding receptor transcript levels during pregnancy . The levels of transcripts for (A) CB1, (B) CB2 and (C) TRPV1 were measured by Q-PCR and normalised against rat β-actin levels. The data were then divided by the mean value obtained on Day 10 and presented as mean ± SEM; n = 4.

Article Snippet: The separated proteins were transferred onto a nitrocellulose membrane in 25 mM Tris-HCl, 250 mM glycine, 20% methanol (v/v), pH 8.3, for 2 h at 200 V. The membranes were incubated with a 1:100 dilution of polyclonal antibodies against CB1 (sc-20754), CB2 (sc-25494) or TRPV1 (sc-12498) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) in blocking solution overnight at 4°C.

Techniques: Binding Assay

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Spatio-temporal expression patterns of anandamide-binding receptors in rat implantation sites: evidence for a role of the endocannabinoid system during the period of placental development

doi: 10.1186/1477-7827-7-121

Figure Lengend Snippet: Anandamide-binding receptor protein levels in mesometrial decidua during pregnancy . (A) Representative western blots for CB1, CB2, TRPV1 and β-tubulin of protein extracts from mesometrial decidua taken on days 10, 12, 14, 16 and 19 of pregnancy. Brain homogenates were used as positive controls (control) for CB1, TRPV1 and spleen homogenates for CB2. β-tubulin was used as a loading control. (B) Densitometric analysis of CB1, CB2 (C) and TRPV1 (D) protein levels. The different protein levels were normalized to the level of β-tubulin. Bar graphs show the mean of experiments carried out with four different animals. Results are depicted as means ± SEM. Data labelled with different letters are significantly different from each other.

Article Snippet: The separated proteins were transferred onto a nitrocellulose membrane in 25 mM Tris-HCl, 250 mM glycine, 20% methanol (v/v), pH 8.3, for 2 h at 200 V. The membranes were incubated with a 1:100 dilution of polyclonal antibodies against CB1 (sc-20754), CB2 (sc-25494) or TRPV1 (sc-12498) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) in blocking solution overnight at 4°C.

Techniques: Binding Assay, Western Blot, Control

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Spatio-temporal expression patterns of anandamide-binding receptors in rat implantation sites: evidence for a role of the endocannabinoid system during the period of placental development

doi: 10.1186/1477-7827-7-121

Figure Lengend Snippet: Immunohistochemical demonstration of anandamide-binding receptor expression in the rat uterus on day 8 of pregnancy . (A) CB1 expression is evenly distributed in the antimesometrial decidua; (B) Higher magnification of the square indicated in (A); (C) CB2 expression in decidual cells of antimesometrium, note that CB2 staining appears to be less intense than CB1; (D) A negative control for CB2 of a consecutive section of (C); (E) TRPV1 expression; note a higher expression in the cells close to the embryo; (F) TRPV1 expression in longitudinal and circular muscle layers. AMD - Antimesometrial decidua; CM - Circular muscle layer; Ec - Embryonic cells; FC - Fibrinoid capsule; LM - Longitudinal muscle layer.

Article Snippet: The separated proteins were transferred onto a nitrocellulose membrane in 25 mM Tris-HCl, 250 mM glycine, 20% methanol (v/v), pH 8.3, for 2 h at 200 V. The membranes were incubated with a 1:100 dilution of polyclonal antibodies against CB1 (sc-20754), CB2 (sc-25494) or TRPV1 (sc-12498) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) in blocking solution overnight at 4°C.

Techniques: Immunohistochemical staining, Binding Assay, Expressing, Staining, Negative Control

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Spatio-temporal expression patterns of anandamide-binding receptors in rat implantation sites: evidence for a role of the endocannabinoid system during the period of placental development

doi: 10.1186/1477-7827-7-121

Figure Lengend Snippet: Spatial distribution of cannabinoid and vanilloid receptors on day 14 of pregnancy . (A) CB1 expression, in the mesometrial decidua and basal zone of placenta; (B) High magnification of the square depicted in (A); note the positive decidual and endothelial cells. (C) Higher magnification image of the basal zone. The giant trophoblast and spongiotrophoblast cells are positive for CB1; (D) Expression of CB2 is lower than CB1 in the mesometrial decidua and basal zone; (E) Negative control for TRPV1; (F) TRPV1 expression in the mesometrial decidua and basal zone; (G) High magnification of the mesometrial decidua and endovascular trophoblast cells; note the lack of staining for TRPV1 in these trophoblast cells and the presence of apoptotic bodies in the decidua (arrowheads); (H) Expression of TRPV1 is particularly evident in uNK cells in the metrial gland region (arrows). BV - Blood vessel; BZ - Basal zone; ET - Endovascular trophoblast cells; GT - Giant trophoblast cells; MD - Mesometrial decidua; MG - Metrial gland; ST - Spongiotrophoblast cells.

Article Snippet: The separated proteins were transferred onto a nitrocellulose membrane in 25 mM Tris-HCl, 250 mM glycine, 20% methanol (v/v), pH 8.3, for 2 h at 200 V. The membranes were incubated with a 1:100 dilution of polyclonal antibodies against CB1 (sc-20754), CB2 (sc-25494) or TRPV1 (sc-12498) from Santa Cruz Biotechnology (Santa Cruz, CA, USA) in blocking solution overnight at 4°C.

Techniques: Expressing, Negative Control, Staining

Journal: Cannabis and Cannabinoid Research

Article Title: Expression of Cannabinoid Receptors in Human Osteoarthritic Cartilage: Implications for Future Therapies

doi: 10.1089/can.2015.0001

Figure Lengend Snippet: Immunohistochemical staining for cannabinoid receptor expression, CB1, CB2, GPR55, GPR18, TRPV1, PPARα, and PPARγ in OA articular cartilage and bone together with rabbit IgG control. CB, cannabinoid; GPR, G-protein-coupled receptor; OA, osteoarthritis; PPAR, peroxisome proliferator-activated receptors; TRPV, transient receptor potential vanilloid.

Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal antibodies (Abcam) against CB1 (1:100) (ab23703), CB2 (1:50)(ab3561), PPARα (1:250)(ab8934), PPARγ (1:50)(ab19481), and TRPV1 (1:1000)(ab63083), and rabbit polyclonal antibodies from Acris (Acris GmbH) against GPR55 (1:100)(SP4239P) and GRP18 (1:200)(AP06930PU).

Techniques: Immunohistochemical staining, Staining, Expressing, Control

Journal: Cannabis and Cannabinoid Research

Article Title: Expression of Cannabinoid Receptors in Human Osteoarthritic Cartilage: Implications for Future Therapies

doi: 10.1089/can.2015.0001

Figure Lengend Snippet: Quantitative analysis of immunopositivity of cannabinoid receptors in each zone of articular cartilage and bone. (A) CB1 immunopositivity. (B) CB2 immunopositivity. (C) GPR55 immunopositivity. (D) GPR18 immunopositivity. (E) TRPV1 immunopositivity. (F) PPARα immunopositivity. (G) PPARγ immunopositivity. * p <0.05, ** p <0.01, *** p <0.001. DZ, deep zone; MZ, middle zone; SZ, superficial zone.

Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal antibodies (Abcam) against CB1 (1:100) (ab23703), CB2 (1:50)(ab3561), PPARα (1:250)(ab8934), PPARγ (1:50)(ab19481), and TRPV1 (1:1000)(ab63083), and rabbit polyclonal antibodies from Acris (Acris GmbH) against GPR55 (1:100)(SP4239P) and GRP18 (1:200)(AP06930PU).

Techniques:

Journal: Cannabis and Cannabinoid Research

Article Title: Expression of Cannabinoid Receptors in Human Osteoarthritic Cartilage: Implications for Future Therapies

doi: 10.1089/can.2015.0001

Figure Lengend Snippet: Quantitative analysis of immunopositivity of cannabinoid receptors in the deep zone of OA articular cartilage. (A) CB1 immunopositivity. (B) CB2 immunopositivity. (C) GPR55 immunopositivity. (D) GPR18 immunopositivity. (E) TRPV1 immunopositivity. (F) PPARα immunopositivity. (G) PPARγ immunopositivity. (H) Correlation of grade of cartilage degeneration and GPR18 immunopositivity. * p <0.05. ID, intermediate degeneration; LD, low degeneration; ND, nondegenerate; SD, severe degeneration.

Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal antibodies (Abcam) against CB1 (1:100) (ab23703), CB2 (1:50)(ab3561), PPARα (1:250)(ab8934), PPARγ (1:50)(ab19481), and TRPV1 (1:1000)(ab63083), and rabbit polyclonal antibodies from Acris (Acris GmbH) against GPR55 (1:100)(SP4239P) and GRP18 (1:200)(AP06930PU).

Techniques:

Journal: Cannabis and Cannabinoid Research

Article Title: Expression of Cannabinoid Receptors in Human Osteoarthritic Cartilage: Implications for Future Therapies

doi: 10.1089/can.2015.0001

Figure Lengend Snippet: Quantitative analysis of immunopositivity of cannabinoid receptors in osteocytes. (A) CB1 immunopositivity. (B) CB2 immunopositivity. (C) GPR55 immunopositivity. (D) GPR18 immunopositivity. (E) TRPV1 immunopositivity. (F) PPARα immunopositivity. (G) PPARγ immunopositivity. (H) Immunopositivity within osteocytes for PPARγ within the underlying bone correlation with grade of cartilage degradation for tissue sample. * p <0.05.

Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal antibodies (Abcam) against CB1 (1:100) (ab23703), CB2 (1:50)(ab3561), PPARα (1:250)(ab8934), PPARγ (1:50)(ab19481), and TRPV1 (1:1000)(ab63083), and rabbit polyclonal antibodies from Acris (Acris GmbH) against GPR55 (1:100)(SP4239P) and GRP18 (1:200)(AP06930PU).

Techniques:

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 1. Immunohistochemistry for CB1 and CB2 in the carotid body. (A) Dot-like CB1 immunoreactivity is observed throughout the carotid body. Intense immu- noreactivity is shown in nerve fibers and/or endings (arrows). (B) A higher magnification view of the rectangle in panel A. Dot-like CB1 immunoreactivity is observed in the perinuclear cytoplasm of chemoreceptor cells (arrows). Intense immunoreactivity for CB1 is also present in nerve endings (arrowheads). (C) Dot-like CB2 immunoreactivity is noted throughout the carotid body. (D) A higher magnification view (rectangle in panel C) shows small dots of CB2 immunoreactivity in the perinuclear region of chemoreceptor cells (arrow). (E-G) Double immunofluorescence for CB1 and CB2 shows CB1-immunoreactive (arrows) and CB2- immunoreactive dots (arrow heads) in the same chemoreceptor cell; however, they are not colocalized.

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunohistochemistry, Immunofluorescence

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 2. (A-C)Triple immunofluorescence for CB1 with TH and DBH. CB1-immunoreactive dot-like structures are observed in both TH- (arrows) and DBH- immunoreactive chemoreceptor cells (arrowheads). Dot-like immunoreactivity for CB1 localizes in both the perinuclear cytoplasm and outlines of the regions of immunoreactivity for TH or DBH. (D-F) Triple immunofluorescence for CB2 with TH and DBH. CB2 immunoreactivity is shown in the perinuclear cytoplasm of TH- (arrows) and DBH-immunoreactive chemoreceptor cells (arrowheads).

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunofluorescence

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 3. (A-C) Double immunofluorescence for CB1 with P2X3. Intense CB1 immunoreactivity is surrounded by P2X3 immunoreactivity in the sensory nerve endings around chemoreceptor cells (arrows). (D-F) Double immunofluorescence for CB1 with VGluT2. CB1 immunoreactivity colocalizes with VGluT2 immunoreactivity, and appears to be in close contact with chemoreceptor cells (arrows). (G-H) Double immunofluorescence for CB2 with P2X2. Weak CB2-immunoreactive dots localize within P2X2-immunoreactive sensory nerve endings (arrows).

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunofluorescence

Journal: World Journal of Gastroenterology

Article Title: Anti-inflammatory effect of cannabinoid agonist WIN55, 212 on mouse experimental colitis is related to inhibition of p38MAPK

doi: 10.3748/wjg.v22.i43.9515

Figure Lengend Snippet: Representative figures of the expression of cannabinoid receptor type 1 and type 2 receptors in mouse colon by immunohistochemistry staining, which are from 4 separate experiments with similar results. A: Expression of cannabinoid (CB)1 receptors; B: Expression of CB2 receptors; (magnification × 400, respectively). The expressions of CB1 receptor in the colonic epithelial cells and CB2 receptor in the submucosal immune cells are marked by arrows and presented as positive results with brown-yellow staining.

Article Snippet: The first antibody was rabbit anti-CB1 receptor or rabbit anti-CB2 receptor polyclonal antibody (1:20 dilution for both; Enzo, Plymouth Meeting, PA, United States).

Techniques: Expressing, Immunohistochemistry, Staining