Journal: Journal of Pain Research

Article Title: Expression of matrix metalloproteinases and components of the endocannabinoid system in the knee joint are associated with biphasic pain progression in a rat model of osteoarthritis

doi: 10.2147/JPR.S132682

Figure Lengend Snippet: Correlation matrix for DWB data and expression levels

Article Snippet: Cannabinoid receptor 2 , Cb2 , Rn04342831_s1 , 847 (NM_020543.4) , 99.

Techniques: Expressing, Gene Expression

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: CB 2 activation reduces the number of infected cells without altering HIV receptor expression. a , b. Bulk RNA-seq analysis showing the expression of key endocannabinoid system genes in ( a ) MDMs and ( b ) iMg. c. Representative images of iMg immunostained for CD4 (red), CCR5 (green), and DAPI (blue). d. Quantification of CD4 and CCR5 expression. Each symbol represents the average of 2 wells/condition. n = 3 differentiations from 2 iMg lines. e. Representative images of iMg immunostained for HIV p24. f. Quantification of CD4 and CCR5 expression. Each symbol represents the average of 2 wells/condition. n = 3 differentiations from 2 iMg lines. One-way analysis of variance (ANOVA) with Dunnett’s correction for multiple comparisons. * p < 0.05; ns, not significant

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: Activation Assay, Infection, Expressing, RNA Sequencing

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: CB 2 agonists dose-dependently reduce HIV replication in macrophages and microglia. a , b. HIV p24 levels in supernatants of monocyte-derived macrophages (MDMs, n = 7) ( a ) and human induced pluripotent stem cell-derived microglia (iMg, n = 3) ( b ), normalized to the HIV + vehicle. Two-way ANOVA with Dunnett’s correction for multiple comparisons. * p < 0.05, ** p < 0.01. c , d. Bulk RNA-seq data showing count/million reads of key endocannabinoid system genes in MDMs ( c ) and iMg ( d )

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: Derivative Assay, RNA Sequencing

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: CB 2 agonists reduce global cytokine release following HIV infection in MDMs but not in iMg. a , b. Cytokine levels in cell-free supernatants of MDM ( a ) and ( b ) iMg cultures were measured with Bio-Plex Pro 48-Plex human cytokine screening panel. Heatmaps show log 2 fold changes from the HIV + vehicle condition. Supernatants were pooled from 4 wells/condition at days postinfection 9 (DPI 9) and assayed in duplicate. n = 6 MDM donors and 3 iMg lines. Student’s t test, ** p < 0.01. c. Levels of cytokines reportedly associated with HIV-induced brain injury in MDM and iMg supernatants, measured with Bio-Plex Pro 48-Plex human cytokine screening panel. d , e. Levels of tumor necrosis factor alpha (TNF-α) and IL-6 in ( d ) MDM and ( e ) iMg culture supernatants, measured with AlphaLISA. n = 6 MDM donors and 3 iMg lines. Student’s t test, * p < 0.05

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: Infection

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: RNA-seq identifies convergent and divergent responses to CB 2 signaling between HIV-infected MDMs and iMg. a , b . Top 10 GO biological process gene sets enriched in the significant differentially expressed genes (DEGs) in comparison between HIV-infected/vehicle-treated and HIV-infected/JWH-133 treated ( a ) MDMs and ( b ) iMg. c. Top 10 KEGG gene sets enriched in the significant DEGs in comparison between HIV-infected/JWH-133-treated MDMs and iMg. n = 2 MDM donors and 2 iMg lines

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: RNA Sequencing, Infection, Comparison

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: CB 2 activation has minimal impact on transcriptional priming of the NLRP3 inflammasome in MDMs and iMg with established HIV infection. (a) NLRP3 inflammasome gene expression in MDMs on DPI 9, determined with qRT-PCR, represented as log 2 fold change from Mock + Veh. n = 3 MDM donors. (b) NLRP3 inflammasome gene expression in iMg on DPI 9, determined with bulk RNA-seq. n = 3 iMg lines

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: Activation Assay, Infection, Gene Expression, Quantitative RT-PCR, RNA Sequencing

Journal: Journal of Neuroimmune Pharmacology

Article Title: Differential Effects of Cannabinoid Receptor 2 Agonists on HIV Replication and Inflammatory Activation in Monocyte-Derived Macrophages and Induced Pluripotent Stem Cell-Derived Microglia

doi: 10.1007/s11481-025-10254-x

Figure Lengend Snippet: In iMg, CB 2 -specific agonists impair HIV-induced NLRP3 inflammasome activation with a corresponding increase in autophagosome number. (a) Quantification of the number of NLRP3 + /ASC + specks in iMg detected with immunostaining. Average of 2 wells/condition from 3 differentiations in 2 iMg lines, one-way ANOVA with Dunnett’s correction for multiple comparisons. * p < 0.05. (b) Autophagic vacuoles measured by an autophagy assay. n = 3 iMg lines, one-way ANOVA with Dunnett’s correction for multiple comparisons, * p < 0.05. (c) Caspase 1 activity measured by the Caspase-Glo 1 inflammasome assay, with results normalized to HIV + Veh. n = 3 iMg lines, one-way ANOVA with Dunnett’s correction for multiple comparisons, * p < 0.05. (d) Levels of IL-1β in iMg supernatants, measured by AlphaLISA. n = 3 iMg lines, one-way ANOVA with Dunnett’s correction for multiple comparisons, ** p < 0.01; ns, not significant

Article Snippet: Custom TaqMan assay plates preloaded with the following human primer/probe pairs (Thermo Fisher Scientific, Waltham, MA) were used: GAPDH , Hs02786624_g1; CD4 , Hs01058407_m1, CCR5 , Hs99999149_s1; CXCR4 , Hs00607978_s1; CNR1 , Hs01038522_s1; CNR2 , Hs00361490_m1; GPR55 , Hs00271662_s1; IL1B , Hs01555410_m1; IL18 , Hs01038788_m1; NLRP3 , Hs00918082_m1; PYCARD , Hs01547324_gH; IL6 , Hs00174131_m1; TNFA , Hs00174128_m1; and CASP1 , Hs00354836_m1.

Techniques: Activation Assay, Immunostaining, Activity Assay

Journal: Scientific Reports

Article Title: Anti-inflammatory effects of CBD in human microglial cell line infected with HIV-1

doi: 10.1038/s41598-023-32927-4

Figure Lengend Snippet: Cannabinoid receptor type 2 gene expression on HC69.5. HIV (GFP+) cells was assessed by RT PCR. H69.5 cells treated with/without 100 μg/ml Poly IC, 1 μM concentration of CBD or /and Δ (9)-THC. After 24 h of treatments, RNAs were extracted, and reverse transcribed followed by real time PCR for CNR2 gene (Hs00275635_m1; Thermofisher Scientific). GAPDH (Hs99999905_m1) was used as a housekeeping gene. Data represent the means ± standard error of three independent experiments. Graph represents the transcript accumulation index respect to negative control C20. Data were analyzed using GraphPad Prism software. The normality distribution was evaluated by Kolmogorov–Smirnov test, while statistical significance was calculated by ANOVA and Tukey Multiple Comparison Test as a post hoc test. Differences were considered significant at p ≤ 0.05.

Article Snippet: After 24 h of treatments, RNAs were extracted, and reverse transcribed followed by real time PCR for CNR2 gene (Hs00275635_m1; Thermofisher Scientific).

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control, Software, Comparison

Journal: Lipids

Article Title: Fatty Acid Binding Protein-1 (FABP1) and the Human FABP1 T94A Variant: Roles in the Endocannabinoid System and Dyslipidemias.

doi: 10.1007/s11745-016-4155-8

Figure Lengend Snippet: Fig. 4 Schematic of FABP1’s role in endocannabinoid (AEA, 2-AG) trafficking and function in primary hepatocytes. By binding ananda- mide (AEA) and 2-arachidonoylglycerol (2-AG), FABP1 may influ- ence key aspects of the hepatic endocannabinoid system: (a) FABP1 may facilitate AEA and 2-AG release/solubilization into the cytosol after their enzymatic cleavage/synthesis from plasma membrane phospholipids by N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD) and diacylglycerol lipase α and β (DAGLα and DAGLβ). FABP1 may or may not facilitate bound AEA and 2-AG for transport/efflux across the plasma membrane and activation of CB receptors on the exofacial leaflet and/or intracellular sites for degra- dation/hydrolysis. FABP1 is known to enhance the cytosolic transport of other bound ligands [5, 46, 52, 141, 251]. (b) FABP1 may facili- tate the reuptake of AEA and 2-AG from the plasma membrane after these lipophilic ligands cross the plasma membrane by diffusion or via G protein-coupled cannabinoid receptor 1 (CB1). Reuptake of 2-AG may also occur via G protein-coupled endocannabinoid recep- tor (CB2). CB2 is expressed only in embryonic liver and in diseased conditions such as fatty liver [252, 253]. CB1 and CB2 activation has been linked to diet-induced hepatic steatosis, primary biliary cir- rhosis, chronic hepatitis, and alcoholic liver [101]. CB1 activation in mice enhances lipogenesis through the sterol regulatory element binding protein-1c (SREBP1c) pathway which induces transcrip- tion of multiple genes in lipogenesis such as SREBP1c itself, acyl CoA carboxylase (ACC), fatty acid synthase (FAS), and stearoyl CoA desaturase (SCD1) [132]. (c) Upon FABP1-mediated release of plasma membrane and/or CB receptor-bound AEA or 2-AG into the cytosol, FABP1 may transport the bound AEA (and possibly 2-AG) into the nucleus for PPARα activation. Although ananda- mide has been shown to bind PPARα and enhance PPARα activation [135], liver fat accumulation in hepatic steatosis results in decreased LCFA oxidation, likely through saturation and/or inhibition of the PPARα pathway by the increasing fat load or damage by inflamma- tion [101]. (d) FABP1 may also transport the bound AEA to the deg- radative enzyme fatty acid amide hydrolase (FAAH) localized with smooth endoplasmic reticulum, mitochondria, lipid droplets, and more rarely at the cell membrane [142–144] by a process analogous to that established for other FABP family members (FABP3, 5, 7) in brain [106–110, 144]. In human liver FABP1 may also transport AEA to/from lysosomes, where it is degraded by N-acylethanolamide acid amide hydrolase (NAAA) [143, 145]. Finally, FABP1 may transport bound 2-AG for degradation by monoacylglycerol lipase (MAGL), an enzyme found at lower levels in liver than brain or other tissues where it is localized diffusely in cytosol and less so in membranes without overall compartmental preference [142]

Article Snippet: ACC Acetyl-CoA carboxylase ACOX1 Acyl-CoA oxidase 1, palmitoyl AEA N-Arachidonoylethanolamide (anandamide) 2-AG 2-Arachidonoylglycerol ALB Albumin ARA C20:4n-6 arachidonic acid CB1 Cannabinoid receptor-1 CB2 Cannabinoid receptor-2 CPT1A Carnitine palmitoyl transferase IA, liver In conjunction with the Lipids 50th Anniversary Symposium, sponsored by Avanti Polar Lipids, at the 106th AOCS Annual Meeting.

Techniques: Binding Assay, Clinical Proteomics, Membrane, Activation Assay, Diffusion-based Assay, Inhibition

Journal: Lipids

Article Title: Fatty Acid Binding Protein-1 (FABP1) and the Human FABP1 T94A Variant: Roles in the Endocannabinoid System and Dyslipidemias.

doi: 10.1007/s11745-016-4155-8

Figure Lengend Snippet: Fig. 6 Human FABP1 T94A variant expression induces transcrip- tion of enzymes and receptors in the endocannabinoid system. Pri- mary human hepatocytes were cultured as described previously [63] followed by determination of mRNA levels encoding the human N-acylphosphatidylethanolamide phospholipase-D (NAPE-PLD), diacylglycerol lipase-α (DAGLα), fatty acid amide hydrolase-1 (FARAH1), fatty acid amide hydrolase (FARAH2), monoacylglyc- erol lipase (MAGL), and cannabinoid receptor-1 (CB1) similarly as for other human mRNA transcripts [213–215]. Values are expressed as the fold change in the ratio of respective mRNA in human FABP1 T94A variant (T94A)/mRNA in wild-type human FABP1. Data are the mean ± SEM (n = 7); *p < 0.05 for T94A vs WT

Article Snippet: ACC Acetyl-CoA carboxylase ACOX1 Acyl-CoA oxidase 1, palmitoyl AEA N-Arachidonoylethanolamide (anandamide) 2-AG 2-Arachidonoylglycerol ALB Albumin ARA C20:4n-6 arachidonic acid CB1 Cannabinoid receptor-1 CB2 Cannabinoid receptor-2 CPT1A Carnitine palmitoyl transferase IA, liver In conjunction with the Lipids 50th Anniversary Symposium, sponsored by Avanti Polar Lipids, at the 106th AOCS Annual Meeting.

Techniques: Variant Assay, Expressing, Cell Culture

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: β‐catenin signalling is inhibited by CB2 gene ablation in d ‐gal‐treated mice. (A) Experimental design. Black bar indicated that mice were administered subcutaneous injections of d ‐gal at 150mg/kg/day for 6 weeks after surgery for 1 week. UNX: unilateral nephrectomy. (B) Representative micrographs showing renal expression of CB2 in different groups. Cryosections were subjected to fluorescence in situ hybridization (FISH) staining for CB2. Arrow indicates positive staining. scale bar, 25 μm. (C) Quantitative real‐time PCR results showing renal expression of CB2. * p < 0.05 versus WT mice group alone; ## p < 0.01 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (D and E) Representative Western blot and quantitative data showing renal expression of β‐catenin. Numbers (1–3) indicate each individual animal in a given group. *** p < 0.001 versus WT mice group alone; ### p < 0.001 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (F and G) Quantitative real‐time PCR results showing renal expression of MMP7 and AT1. * p < 0.05 and *** p < 0.001 versus WT mice group alone; # p < 0.05 and ## p < 0.01 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (H) Representative micrographs showing the expression of active β‐catenin. Frozen kidney sections were stained with an antibody against active β‐catenin. Arrow indicates positive staining. Scale bar, 75μm. (I) Quantitative data showing quantification of positive staining. * p < 0.05 versus WT mice group alone; # p < 0.05 versus the d ‐gal‐treated WT mice group alone (n = 5–6)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Expressing, Fluorescence, In Situ Hybridization, Staining, Real-time Polymerase Chain Reaction, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 is upregulated in aged kidneys. (A) Representative micrographs showing CB2 expression in kidneys from 2‐month‐old and 24‐month‐old mice. Cryosections were subjected to fluorescence in situ hybridization (FISH) staining for CB2. Arrow indicates positive staining. scale bar, 25 μm. (B‐E) Representative Western blot and quantitative data showing renal expression of CB2 from 2‐month‐old and 24‐month‐old mice (B and C) or mice which were administered subcutaneous injections of d ‐gal at 150mg/kg/day for 6 weeks (D and E). Numbers (1–5) indicate each individual animal in a given group. ** p < 0.01 versus 2‐month‐old mice group or the sham control group (n = 5). (F) Representative images showing renal expression of CB2 in d ‐gal‐treated mice. Cryosections were subjected to fluorescence in situ hybridization (FISH) staining for CB2. Arrow indicates positive staining. scale bar, 25 μm. (G) Representative micrographs showing the colocalization of CB2 and various segment‐specific tubular markers in kidneys. Frozen kidney sections were stained for CB2 (red) using FISH and various segment‐specific tubular markers (green) by immunofluorescence. The following segment‐specific tubular markers were used: proximal tubule, lotus tetragonolobus lectin (LTL); distal tubule, peanut agglutinin (PNA); arrows indicate positive tubules with colocalization of CB2 and specific tubular markers. Scale bar, 25 μm. (H) Representative micrographs showing the expression of CB2 and TOMM20 in tubules in 2‐month‐old and 24‐month‐old mice. Cryosections were subjected to FISH staining of CB2 (red) and stained with TOMM20 (green) antibody by immunofluorescence. Arrows indicate positive staining. Scale bar, 25μm

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Expressing, Fluorescence, In Situ Hybridization, Staining, Western Blot, Control, Immunofluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 gene ablation does not affect kidney ageing or mitochondrial function in young mice. (A) RT‐PCR analyses showing renal expression of CB2 in wild‐type mice (WT) and CB2 knockout mice (KO). Numbers (1–4) indicate each individual animal in a given group. (B‐F) Quantitative real‐time PCR results showing renal expression of CB2, fibronectin, α‐SMA, CollagenⅠa1 and CollagenⅢa1 in WT and KO mice. ** p < 0.01, n.s. versus WT mice group (n = 4). n.s.: no significance. (G) Representative micrographs showing Periodic acid‐Schiff (PAS) staining, Sirius red staining, senescence‐associated β‐galactosidase activity (SA‐β‐gal) staining and the expression of TOMM20. Paraffin‐embedded kidney sections were subjected to PAS and Sirius red staining. Frozen kidney sections were stained for SA‐β‐gal activity and TOMM20. Scale bar, 50 μm. (H‐K) Representative Western blot and quantitative data showing renal expression of PGC‐1α, TOMM20 and TFAM in WT and KO mice. Numbers (1–4) indicate each individual animal in a given group. n.s. versus WT mice group (n=4). n.s.: no significance. (L‐M) Quantitative real‐time PCR results showing renal expression of p16 INK4A and γH2AX in WT and KO mice. n.s. versus WT mice group (n = 4). n.s.: no significance. (N‐R) Representative Western blot and quantitative data showing renal expression of β‐catenin, MMP7, Snail1 and AT1 in WT and KO mice. Numbers (1–4) indicate each individual animal in a given group. n.s. versus WT mice group (n = 4). n.s.: no significance

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Knock-Out, Real-time Polymerase Chain Reaction, Staining, Activity Assay, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 deficiency protects renal mitochondrial homeostasis in the accelerated ageing mice. (A) Representative micrographs showing renal expression of PGC‐1α and TOMM20 in different groups. Paraffin‐embedded kidney sections were immunostained with an antibody against PGC‐1α or TOMM20. Arrows indicate positive staining. Scale bar, 50 μm. (B‐C) Quantitative data showing quantification of positive staining. * p < 0.05, *** p < 0.001 versus WT mice group alone; # p < 0.05, ### p < 0.001 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (D) Representative graph showing the production of adenosine triphosphate (ATP) in different groups. * p < 0.05 versus WT mice group alone; ## p < 0.01 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (E–H) Representative Western blot and quantitative data showing renal expression of PGC‐1α, TOMM20 and Cytb. Numbers (1–3) indicate each individual animal in a given group. * p < 0.05, ** p < 0.01, *** p < 0.001 versus the WT mice group alone; # p < 0.05, ### p < 0.001 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (I) Representative transmission electron microscopy graphs showing mitochondrial ultrastructure of renal tubular cells in different groups. Arrows indicate damaged and abnormal‐shaped mitochondria. Scale bar, 1μm

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Expressing, Staining, Western Blot, Transmission Assay, Electron Microscopy

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 gene ablation ameliorates kidney ageing. (A) Representative micrographs showing renal expression of γH2AX and SA‐β‐gal activity in different groups. Paraffin‐embedded kidney sections were immunostained with an antibody against γH2AX (top). Frozen kidney sections were stained for SA‐β‐gal activity (bottom). Arrows indicate positive staining. Scale bar, 50 μm. (B‐C) Quantitative data showing quantification of positive staining. ** p < 0.01 versus WT mice group alone; ## p < 0.01 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (D–G) Representative Western blot and quantitative data showing renal expression of p16 INK4A , γH2AX and p19 ARF in different groups. Numbers (1–3) indicate each individual animal in a given group. ** p < 0.01, *** p < 0.001 versus the WT mice group alone; # p < 0.05, ## p < 0.01, ### p < 0.001 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (H and I) Representative micrographs showing renal expression of klotho in different groups (H). Paraffin‐embedded kidney sections were immunostained with an antibody against klotho. Arrows indicate positive staining. Scale bar, 50 μm. (I) Quantitative data showing quantification of positive staining. *** p < 0.001 versus the WT mice group alone; ### p < 0.001 versus the d ‐gal‐treated WT mice group alone (n = 5–6)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Expressing, Activity Assay, Staining, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 deficiency retards age‐related kidney fibrosis. (A and B) Quantitative data showing serum creatinine (Scr) and blood urea nitrogen (BUN) levels in different groups. n.s.: no significance. (C–E) Representative Western blot and quantitative data showing renal expression of fibronectin and α‐SMA in different groups. Numbers (1–3) indicate each individual animal in a given group. * p < 0.05 versus the WT mice group alone; # p < 0.05, ## p < 0.01 versus the d ‐gal‐treated WT mice group alone (n = 5–6). (F–H) Representative micrographs showing renal expression of fibronectin and Sirius red staining in different groups. Paraffin‐embedded kidney sections were stained with Sirius red and were immunostained with an antibody against fibronectin. Arrows indicate positive staining. Scale bar, 50 μm. Quantitative data showing quantification of positive staining of fibronectin (G) and fibrotic area (H). ** p < 0.01, *** p < 0.001 versus the WT mice group alone; ## p < 0.01, ### p < 0.001versus the d ‐gal‐treated WT mice group alone (n = 5–6)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Western Blot, Expressing, Staining

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 induces mitochondrial dysfunction and cellular senescence in vitro. (A–I) Representative Western blot and quantitative data showing the expression of CB2, PGC‐1α, Cytb, TOMM20, COX1, COX2, p16 INK4A , γH2AX in HKC‐8 cells. HKC‐8 cells were transfected with CB2 expression plasmid (pCMV‐CB2) for 24 h. * p < 0.05, ** p < 0.01 versus the pcDNA3 group (n = 3). (J–T) Representative Western blot and quantitative data showing the expression of CB2, PGC‐1α, Cytb, TOMM20, COX1, COX2, TFAM, p16 INK4A , γH2AX and p14 ARF in HKC‐8 cells. HKC‐8 cells were treated with AM1241 (10 μM) for 48h. * p < 0.05, ** p < 0.01 versus the control group (n = 3)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: In Vitro, Western Blot, Expressing, Transfection, Plasmid Preparation, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: β‐catenin plays a mediative role in CB2‐induced mitochondrial dysfunction and cellular senescence. (A–E) Representative Western blot (A, F) and quantitative data (C–E, G and H) showing the expression of COX1, TFAM, TOMM20, p14 ARF and γH2AX in HKC‐8 cells. HKC‐8 cells were treated with AM1241 (10 μM) for 48 h and pretreated with XL‐001 (10 μM) for 1 h. Quantitative data graph (B) showing the production of adenosine triphosphate (ATP) in HKC‐8 cells. * p < 0.05, ** p < 0.01, *** p < 0.001 versus the control group alone; # p < 0.05, ## p < 0.01, ### p < 0.001versus the AM1241 group alone (n = 3). (I–N) Representative Western blot (I, M) and quantitative data (J–L, N) showing the expression of PGC‐1α, Cytb, TOMM20 and p16 INK4A in HKC‐8 cells. HKC‐8 cells were transfected with CB2 expression plasmid (pCMV‐CB2), followed by the stimulation of ICG‐001 at 10μM for 24 h * p < 0.05, ** p < 0.01, *** p < 0.001 versus the control group alone; # p < 0.05, ## p < 0.01versus the pCMV‐CB2 group alone (n = 3)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Western Blot, Expressing, Control, Transfection, Plasmid Preparation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Cannabinoid receptor 2 plays a central role in renal tubular mitochondrial dysfunction and kidney ageing

doi: 10.1111/jcmm.16857

Figure Lengend Snippet: CB2 plays a central role in the accelerated ageing in renal tubular cells. (A–H) Representative Western blot and quantitative data showing the expression of CB2, PGC‐1α, TOMM20, COX1, p16 INK4A , p14 ARF and β‐catenin in HKC‐8 cells. HKC‐8 cells were treated with D‐gal at 10mg/ml for 72h and pretreated with XL‐001 (10μM) for 1 h. * p < 0.05, ** p < 0.01, *** p < 0.001 versus the control group alone; # p < 0.05, ## p < 0.01, ### p < 0.001 versus the d ‐gal group alone (n = 3). (I and J) Representative micrographs and quantitative data showing SA‐β‐gal activity in different groups. Frozen kidney sections were stained for SA‐β‐gal activity. Arrows indicate positive staining. Scale bar, 20 μm. *** p < 0.001 versus the control group alone; ### p < 0.001 versus the d ‐gal group alone (n = 3). (K–N) Representative Western blot and quantitative data showing renal expression of PGC‐1α, TOMM20 and p14 ARF in HKC‐8 cells. HKC‐8 cells were treated with D‐gal at 10mg/ml or cotreated with AM1241 (10 μM) for 72 h and pretreated with ICG‐001 (10 μM) for 1 h. * p < 0.05, ** p < 0.01, *** p < 0.001 versus the control group alone; # p < 0.05, ## p < 0.01, ### p < 0.001 versus the d ‐gal group alone; †† p < 0.01, ††† p < 0.001 versus the d ‐gal+AM1241 group alone (n = 3). (O and P) Representative micrographs and quantitative data showing SA‐β‐gal activity in different groups. Frozen kidney sections were stained for SA‐β‐gal activity. Arrows indicate positive staining. Scale bar, 20 μm. ** p < 0.01 versus the control group alone; ### p < 0.001 versus the d ‐gal group alone; ††† p < 0.001 versus the d ‐gal+AM1241 group alone (n = 3)

Article Snippet: The expression of CB2 was assayed by fluorescence staining using a Fluorescence in situ hybridization kit (MK2530‐m; Boster technology).

Techniques: Western Blot, Expressing, Control, Activity Assay, Staining

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: List of primers used to determine the relative expression of canonical and extended endocannabinoid receptors in mice ( Mus musculus ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing, Mouse Assay

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: List of primers used to determine the relative expression of canonical and extended endocannabinoid receptors in rats ( Ratus norvegicus ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: List of primers used to determine the relative expression of canonical and extended endocannabinoid receptors in Rhesus macaques ( Macaca mulatta ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: Both Canonical ECS Receptors Are Present In The Spleen Of Mice, Rats, and NHP. Relative expression of cnr1 and cnr2 was determined using qPCR from the colon, heart, kidney, liver, MLN, spleen, and visceral fat from mice, rats, and NHP. (a) In mice, we detected cnr1 mRNA in the colon, kidney (4/5 mice), liver (2/5 mice), spleen, and visceral fat, having the highest levels in the latter. Significant differences were found in the levels of expression between the colon and the heart, as well as among the visceral fat and the levels of expression in the heart and the liver. (b) C nr1 was detected in partial samples of the rat model colon (4/6 rats), heart (1/6 rats), kidney, and spleen (1/6 rats), having statistically higher levels of expression in the kidney when compared to the heart, liver, MLN, and spleen. (c) In NHP, cnr1 mRNA was detected in the spleen (2/3 rhesus) and visceral fat at comparable levels. (d) Cnr2 was detected only in the spleen and visceral fat (4/5 mice), at comparable levels. The spleen had statistically significantly higher levels as compared to the colon, heart, kidney, and liver. (e) Cnr2 was detected in the colon (2/4 rats), heart (1/6 rats), MLN (4/6 rats), and spleen of rats. (f) In NHP, cnr2 mRNA was detected in the liver (1/4 rhesus), MLN, and spleen. Detection levels were significantly higher in the spleen when compared to the heart, kidney, and liver, but with significant differences when compared to the liver (1/4 rhesus), kidney, and heart. Data are represented as the mean ± SD.

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: Summary of the findings of the expression of the canonical and extended ECS in mice ( Mus musculus ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing, Mouse Assay

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: Summary of the findings of the expression of the canonical and extended ECS in rats ( Ratus norvegicus ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing

Journal: Physiological Reports

Article Title: Roadmap for the expression of canonical and extended endocannabinoid system receptors and metabolic enzymes in peripheral organs of preclinical animal models

doi: 10.14814/phy2.15947

Figure Lengend Snippet: Summary of the findings of the expression of the canonical and extended ECS in Rhesus macaques ( Macaca mulatta ).

Article Snippet: htr1a , Rn01637601_m1 , ThermoFisher Scientific.

Techniques: Expressing