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: The FASEB Journal

Article Title: Inhibition of FAAH suppresses RANKL ‐induced osteoclastogenesis and attenuates ovariectomy‐induced bone loss partially through repressing the IL17 pathway

doi: 10.1096/fj.202200911R

Figure Lengend Snippet: PF inhibits osteoclast differentiation – CB1 or CB2 independence. (A–C) BMMs were infected with the lentivirus carrying FAAH‐specific shRNA or the control lentivirus and then treated with RANKL for 3 days, and the mRNA and protein expression of CB1 and CB2 were detected by qPCR and Western blot respectively. (D–F) BMMs were treated with indicated concentrations of PF (0, 0.5, 1, and 2.5 μM) with or without stimulation by 75 ng/ml RANKL for 3 days, and the mRNA and protein expression of CB1 or CB2 were determined using qPCR and Western blot respectively. # p < .05, ## p < .01, compared with the control groups; ns, non‐significant, compared with the RANKL groups. (G and H) BMMs were treated with PF (2.5 μM), AM251 (0.1 μM), and AM630 (0.1 μM) as indicated in the presence of 75 ng/ml RANKL. After 5–7 d, trap staining was conducted, and multinucleated (>3 nuclei) cells were quantified (scale bar = 400 μm). (I–K) BMMs were treated with PF (2.5 μM), AM251 (0.1 μM), and AM630 (0.1 μM) as indicated in the presence of 75 ng/ml RANKL for 3 days, and the mRNA and protein expression of osteoclast‐specific genes were determined using qPCR and Western blot respectively. # p < .05, ## p < .01, compared with the RANKL groups; ns, non‐significant, compared with the RANKL + PF groups. All data are presented as the mean ± SD. N = 3 per group.

Article Snippet: Primary antibodies targeting FAAH, CB1, and CB2 were purchased from Abclonal (Wuhan, Hubei, China).

Techniques: Infection, shRNA, Control, Expressing, Western Blot, Staining

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: Journal of Neuroimmune Pharmacology

Article Title: Alleviating CB2-Dependent ER Stress and Mitochondrial Dysfunction Improves Chronic Cerebral Hypoperfusion-Induced Cognitive Impairment

doi: 10.1007/s11481-024-10098-x

Figure Lengend Snippet: Effects of URB597 and 4-PBA on the CB2/β-Arrestin1-dependent ER stress and mitochondrial apoptosis. A Immunoprecipitation confirmed the interaction between CB2 and β-Arrestin1. B Representative western blots of CB2, β-Arrestin1, p-PERK/PERK, CHOP, Cyt-c, and caspase-9. C – H Quantitative analysis of protein expression. * p < 0.05 vs. Sham; # p < 0.05 νs. M, ^ p < 0.05 νs. MP, & p < 0.05 νs. MU, ▽ p < 0.05 νs. MUP ( n = 3)

Article Snippet: The FAAH inhibitor (URB597) (No. HY-10,864), the ER stress inhibitor and inducer, [benzenebutyric acid (4-PBA) (No. HY-A0281) and thapsigargin (TG) (No. HY-13,433), respectively], and a selective CB2 antagonist (6-lodopravadoline, AM630) (No. HY-15,421) were all purchased from the Med Chem Express (MCE, Shanghai, China).

Techniques: Immunoprecipitation, Western Blot, Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Update on the controversial identity of cells expressing cnr2 gene in the nervous system

doi: 10.1111/cns.13977

Figure Lengend Snippet: List of anti‐CB2R antibodies used for the detection of CB2R protein in the central nervous system

Article Snippet: D , #ACR‐002 , Alomone , Yes , aa 228–242 of rat CB2R , Rabbit , Poly , Mouse, rat.

Techniques:

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