Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Expression of cannabinoid and cannabinoid-like receptors in brain endothelial and melanoma cells. RT-PCR was performed to determine the expression of CB2A and CB2B (positive control: HL-60) transcriptional variants of human CB2 receptor in hCMEC/D3 brain endothelial and A2058 melanoma cells ( a , b ); the expression of transcriptional variant 1 and 2 of rat CB2 receptor ( c , d ) in rat brain endothelial cells (RBECs) (positive control: rat spleen), the expression of CB1 receptor ( e ); transcriptional variant 1 and 2 of GPR18 ( f , g ); GPR119 ( h ) and GPR55 ( i ) in hCMEC/D3 human brain endothelial cells and A2058 melanoma cells. Dotted arrows indicate the absence of specific bands.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Variant Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the attachment of melanoma cells on the brain endothelium. Results are represented as % control ( i.e. , D3 + A2058) and given as mean ± SD. N = 3. * p < 0.05 as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) D3(jwh-133) and A2058(jwh-133) represent cells pre-treated with 10 μM JWH-133 for 4 h. D3 + A2058 + JWH-133 denotes cells treated with 10 μM JWH-133 during the 90 min adhesion assay; ( b ) JWH-133 (10 μM), U0126 (10 μM) and PTX (100 ng/mL) were applied in pre-treatment of both cell types and treatment during the adhesion assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Control, Cell Adhesion Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the transendothelial migration of melanoma cells. Results are represented as % control ( i.e. , RBEC + A2058) and given as mean ± SD. N = 3. * p < 0.01 (compared to control) as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) RBEC(jwh-133) represents endothelial cells pre-treated with 10 μM JWH-133 for 4 h. RBEC(jwh-133) + A2058(jwh-133) + JWH-133 denotes that both endothelial and melanoma cells were pre-treated with 10 μM JWH-133 for 4 h and treated with 10 μM JWH-133 during the 5 h transmigration assay; ( b ) JWH-133 (10 μM) and SR-144528 (10 μM) were applied in pre-treatment of both cell types and treatment during the transmigration assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Migration, Control, Transmigration Assay

Journal: Nature neuroscience

Article Title: Brain cannabinoid CB₂ receptors modulate cocaine's actions in mice.

doi: 10.1038/nn.2874

Figure Lengend Snippet: Figure 1 Effects of JWH133 on cocaine self-administration. (a) Systemic administration of JWH133 (10 and 20 mg per kg, intraperitoneal, 30 min before testing) inhibited cocaine self-administration under FR1 reinforcement in wild-type (one-way ANOVA, F2,16 = 13.09, P < 0.001) and CB1

Article Snippet: JWH133, AM251 and AM630 were obtained from Tocris Bioscience.

Techniques:

Journal: Nature neuroscience

Article Title: Brain cannabinoid CB₂ receptors modulate cocaine's actions in mice.

doi: 10.1038/nn.2874

Figure Lengend Snippet: Figure 3 Systemic administration of JWH133 (10 and 20 mg per kg, intraperitoneal, 30 min before cocaine) dose-dependently inhibited cocaine-enhanced locomotion in wild-type (a, two-way ANOVA for repeated measures over time, F2,16 = 14.45, P < 0.001) and CB1

Article Snippet: JWH133, AM251 and AM630 were obtained from Tocris Bioscience.

Techniques:

Journal: Nature neuroscience

Article Title: Brain cannabinoid CB₂ receptors modulate cocaine's actions in mice.

doi: 10.1038/nn.2874

Figure Lengend Snippet: Figure 5 Effects of systemic JWH133 and/or AM630 on NAc DA. (a–f) Systemic administration of JWH133 (3, 10 or 20 mg per kg, intraperitoneal) dose-dependently inhibited basal (a–c) or cocaine-enhanced (d–f) extracellular NAc DA in wild-type (a, two-way ANOVA for repeated measures over time, F3,29 = 25.97, P < 0.001; d, F2,19 = 4.47, P < 0.05) and CB1

Article Snippet: JWH133, AM251 and AM630 were obtained from Tocris Bioscience.

Techniques:

Journal: Scientific Reports

Article Title: The Cannabinoid Receptor 2 Protects Against Alcoholic Liver Disease Via a Macrophage Autophagy-Dependent Pathway

doi: 10.1038/srep28806

Figure Lengend Snippet: ( A ) Representative images of CB2 (red) and F4/80 (green) labeling in peritoneal macrophages isolated from WT and CB2 Mye −/− mice (original magnification x400). ( B ) mRNA expression of CCL3, IL-6, IL-1β, IL-1α and TNF-α in Kupffer cells isolated from WT and CB2 Mye −/− mice and exposed to 1 ng/ml of LPS for 6 hours. Data are mean ± SEM of 5–10 samples per condition. & p < 0.05 for WT vs CB2 Mye −/− mice.

Article Snippet: The CB2 agonist JWH-133 was obtained from Axon Medchem, absolute ethanol from Carlo Erba Reactifs and Brewer Thioglycholate medium from BD Pharmingen.

Techniques: Labeling, Isolation, Expressing

Journal: Scientific Reports

Article Title: The Cannabinoid Receptor 2 Protects Against Alcoholic Liver Disease Via a Macrophage Autophagy-Dependent Pathway

doi: 10.1038/srep28806

Figure Lengend Snippet: ( A ) Hepatic mRNA expression of CCL3, IL-6, IL-1β, IL-1α and TNF-α in control diet (CD)- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice. ( B ) Representative images (original magnification x400) and quantification of F4/80 staining in CD- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice. ( C ) Representative images (original magnification x200) and quantification of MPO staining in CD- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice. Arrows indicate positive cells. ( D ) Hepatic mRNA expression of Ly6G, SELE, SELP, ICAM1, ESL-1 and CD44 in CD- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice. ( E ) Left , representative hematoxylin-eosin staining of liver tissue sections from CD- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice (original magnification x200), and right , hepatic triglycerides content of CD- and chronic-plus-binge ethanol-fed WT and CB2 Mye −/− mice. Data are mean ± SEM. *p < 0.05 for CD vs ethanol and & p < 0.05 for WT vs CB2 Mye −/− mice.

Article Snippet: The CB2 agonist JWH-133 was obtained from Axon Medchem, absolute ethanol from Carlo Erba Reactifs and Brewer Thioglycholate medium from BD Pharmingen.

Techniques: Expressing, Control, Staining

Journal: Scientific Reports

Article Title: The Cannabinoid Receptor 2 Protects Against Alcoholic Liver Disease Via a Macrophage Autophagy-Dependent Pathway

doi: 10.1038/srep28806

Figure Lengend Snippet: Cells were exposed to 5 μM of JWH-133, 100 nM rapamycin or vehicle for 6 hours in the presence or absence of 10 μM of chloroquine (CQ). ( A ) Western blot analysis and quantification of LC3, SQSTM1/p62 and β-actin protein expression in RAW264.7 cells. ( B ) Representative images (original magnification x400) and quantification of the number of LC3-positive dots ( left ) and SQSTM1/p62-positive dots ( right ) in RAW264.7 cells. ( C ) Representative images (original magnification x400) and quantification of the number of GFP-LC3-positive dots in F4/80 (red)-positive peritoneal macrophages ( left ) and SQSTM1/p62 (red)-positive dots in F4/80 (green)-positive peritoneal macrophages ( right ) isolated from GFP-LC3 transgenic mice. ( D ) Representative images (original magnification x400) and quantification of LC3-positive dots in RAW264.7 cells transfected with the RFP-GFP-LC3 plasmid (autophagosomes = GFP + RFP + (yellow dots), autolysosomes = GFP - RFP + (red dots), Total = GFP + RFP + and GFP - RFP + ). ( E ) Representative images (original magnification x400) and quantification of the number of LC3 (red)-positive dots ( left ) and of SQSTM1/p62 (red)-positive dots ( right ) in F4/80 (green)-positive peritoneal macrophages isolated from WT and CB2 Mye −/− mice and exposed to 10 μM of chloroquine or its vehicle for 6 hours. Data are mean ± SEM. # p < 0.05 for JWH-133 or rapamycin vs vehicle, * p < 0.05 for - CQ vs +CQ, & p < 0.05 for WT vs CB2 −/− cells.

Article Snippet: The CB2 agonist JWH-133 was obtained from Axon Medchem, absolute ethanol from Carlo Erba Reactifs and Brewer Thioglycholate medium from BD Pharmingen.

Techniques: Western Blot, Expressing, Isolation, Transgenic Assay, Transfection, Plasmid Preparation