Journal: Journal of Cannabis Research

Article Title: Cannabidiol does not cause DNA double-strand breaks in a human liver-derived cell model

doi: 10.1186/s42238-025-00365-w

Figure Lengend Snippet: HepG2 cells show a concentration-dependent reduced expression of metabolic, DNA repair and uncleaved apoptotic proteins. A Immunoblot using antibodies against cannabinoid-receptor (CB) 1, CB 2, caspase-3 and LEDGF. Cells were incubated for 24 h with 5 - 50 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mu M$$\end{document} CBD. GAPDH was used as loading control and the whole protein amount was verified using Coomassie brilliant blue staining. No expression of CB1 was observed. Expression of CB2 in HepG2 cells was confirmed by observing a 52/55 kDA protein band (receptor doublet). B - D Quantitative analysis of CB2, caspase-3 and LEDGF/p75 expression ( \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$n = 3$$\end{document} ). E Analysis of dose-dependent cAMP concentration after increasing concentrations of CBD. \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$p < 0.05 = *$$\end{document} , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$p < 0.01 = **$$\end{document} , \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$p < 0.005 = ***$$\end{document}

Article Snippet: The membrane was incubated with an solution (antibody diluted in 2 % non-fat dried milk in TBST) containing the primary antibodies against LEDGF/p75((Bethyl laboratories, Cat# A300-847A, RRID: AB 609466, 1:1000 dilution), CB-1 (Santa Cruz Biotechnology, Dallas, USA, Cat# sc-293419, RRID: N/A, 1:1000 dilution), CB-2 (Santa Cruz Biotechnology, Dallas, USA, Cat# sc-293188, RRID: N/A), GAPDH (Cell Signaling, Technology, Massachusetts, USA, Cat# 2118, RRID: AB 561053, 1: 20.000) and Caspase-3 (Cell Signaling technology, Messacchusetty, USA, Cat# 9662, RRID: AB 331439, 1:1000) for 1 hour at room temperature.

Techniques: Concentration Assay, Expressing, Western Blot, Incubation, Control, Staining

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: HIV-infected MDM keep expressing surface CB2R at day 12pi. MDM were labeled with a fluorescent CB2R monoclonal antibody (FL4 channel) conjugated with Alexa Fluor® 647 and then analyzed by flow cytometry. ( a ) Representative histograms of extracellular staining show significant differences in fluorescence intensities from HIV + (dark histogram) and uninfected (grey histogram) on the expression of CB2R compared with the nonfluorescent MDM (unshaded histograms to the left. ( b ) A graphic representation of surface CB2R levels per donor is shown. Panels a and b are representative of three different donors (n = 3). An unstained control was included as a technical negative control and is representative of 1 donor (n = 1). ( c ) Representative images of intracellular levels of CB2R and GAPDH expression using Western Blot. Images were cropped from the same blot after stripping and reprobing with a different antibody and were acquired using different exposure times. Full-length blots are presented in Supplementary Fig. 4 ( d ) A graphic representation of intracellular CB2R levels per donor is shown. Figures c and d are representative of four different donors (n = 4).

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Infection, Expressing, Labeling, Flow Cytometry, Staining, Fluorescence, Control, Negative Control, Western Blot, Stripping Membranes

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: CB2R agonists treatment decrease HIV-1 replication and CATB secretion from MDM. MDM were infected with HIV-1 and treated with CB2R agonists, JWH-133 and HU-308. HIV-1 p24 and CATB levels were measured from supernatants of days 3, 6, 9, and 12pi by ELISA. ( a ) Temporal HIV-1 p24 levels in supernatants of HIV-infected MDM treated with JWH-133. ( b ) Temporal CATB secretion levels in supernatants of HIV-infected MDM treated with JWH-133. Figures a and b are representative of at least four different donors (n = at least 4). ( c ) Temporal HIV-1 p24 levels in supernatants of HIV-infected MDM treated with HU-308. ( d ) Temporal CATB secretion levels in supernatants of HIV-infected MDM treated with HU-308. Figures c and d are representative of at least six different donors (n = at least 6). Graphs are presented using the mean and the ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01 vs. vehicle control at its respective time-point.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Control

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: JWH-133 prevents HIV-induced increase in surface CB2R expression and induces oscillating expressions over time. MDM were cultured in 8-well chamber slides, infected with HIV-1 ADA , and treated with JWH-133 at 0.5 µM. Slides were fixed at days 3, 6, 9, and 12dpi. Cells were stained with an anti-CB2R antibody (red) and nuclei were stained with DAPI (blue). At least 3 different random pictures per condition were acquired. Representative immunofluorescence images are shown. Graphs are presented using the mean ± standard error of the mean (SEM). This figure is representative of three different donors (n = 3). * p < 0.05, ** p < 0.01.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Expressing, Cell Culture, Infection, Staining, Immunofluorescence

Journal: Scientific Reports

Article Title: Cannabinoid receptor type 2 agonist JWH-133 decreases cathepsin B secretion and neurotoxicity from HIV-infected macrophages

doi: 10.1038/s41598-021-03896-3

Figure Lengend Snippet: JWH-133 decreases HIV-1 replication and CATB secretion through CB2R activation. MDM were cultured in 24-wells plates and infected with HIV-1 ADA . After removal of residual virus, cells were treated with CB2R antagonist SR144528 (SR: 1 µM) for 1 h, followed by JWH-133 (JWH) treatment at 0.5 µM. Treatments were maintained for 6dpi, exchanging half of the media at day 3pi and repeating the co-administration protocol. HIV-1 p24 and CATB levels were measured in HIV-infected MDM supernatants using ELISA. HIV-1 p24 and CATB levels were normalized against its vehicle control per donor. ( a ) HIV-1 p24 levels in HIV-infected MDM after co-administration of CB2R ligands. This figure is representative of at least eight different donors (n = 8). ( b ) CATB levels in HIV-infected MDM after co-administration of CB2R ligands. This figure is representative of ten different donors (n = 10). Graphs are presented using the mean ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01.

Article Snippet: MDM were incubated with an anti-human CB2R-Alexa Fluor® 647 conjugated antibody (R&D Systems) (1:100 in PBS 1X) for 1 h at 4 °C.

Techniques: Activation Assay, Cell Culture, Infection, Virus, Enzyme-linked Immunosorbent Assay, Control

Journal: Annals of Dermatology

Article Title: Selective Cannabinoid Receptor-1 Agonists Regulate Mast Cell Activation in an Oxazolone-Induced Atopic Dermatitis Model

doi: 10.5021/ad.2016.28.1.22

Figure Lengend Snippet: Newly synthesized cannabinoids and the cAMP assay screening for cannabinoid receptor modulators. (A) The structure of α-oleoyl oleoylamine ethanolamine (α-OOE) and α-oleoyl oleoylamine serinol (α-OOS). (B, C) Synthetic compounds were screened in stable CHO cell lines overexpressing cannabinoid receptor-1 (CB1R) or CB2R. Pre-treatment with CBR agonists significantly diminished the cytosolic levels of cAMP induced by 10 µM forskolin (B). N-arachidonoylethanolamine (AEA) and Hu308 were used as controls for CB1R and CB2R, respectively. All data are presented as the mean±standard error, t -test and analysis of variance. * p <0.01.

Article Snippet: To establish stable cell lines expressing CBR, CHO-K1 cells were transfected with human CB1R (Cat. RC210397) and CB2R (Cat. SC118984) cDNA constructs (Origene, Rockville, MD, USA); the following day, media were replaced with F12 standard medium containing G418 (100 μg/ml) to select for stable clones.

Techniques: Synthesized, cAMP Assay

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