Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Induction of the endocannabinoid system in acute pancreatitis. Immunohistochemical detection of CB1 and CB2 is shown in human pancreas (A–C and G–I) or mouse pancreas (D–F and J–L) derived either from control subjects (A, D, G, and J) or subjects with acute pancreatitis (B and C, E and F, H and I, K and L). (C, F, I and L) Areas of necrosis within human and mouse acute pancreatitis are indicated by dotted lines. In normal human pancreas (A) CB1-immunoreactivity in acinar cells (▶; magnified in inset C), ducts (→), and nerves (magnified in insets), was weak but strong in acute pancreatitis (B, C). (D–F) Similarly, pancreas demonstrated increased immunoreactivity for CB1 after induction of acute pancreatitis with cerulein (E, F) over normal mouse pancreas (D). Intense CB1 immunostaining is observed on acinar cell necrosis (dotted arrow; F). (G–I) Moderate staining for CB2 is seen in normal human pancreas (G) with a slight increase in acute pancreatitis (H, I) in acinar cells (▶; magnified in inset I), ducts (→), and nerves (magnified in insets). Upon induction of acute pancreatitis in mice, there is a pronounced increase in CB2 immunoreactivity over expression levels in normal mouse pancreas (J–L). This is particularly present within acinar cell necrosis (dotted arrow; L). Original magnification: 40x (A and B, D and E, G and H, J and K) or 80x (F and L; insets in A–C and G–I) objective. (M, N) Immunoblot analysis of pancreas samples derived from acute pancreatitis (lanes 1–3) and control human donors (lanes 4–6) with antibodies recognizing CB1 (M) or CB2 (N). (▶) indicates anti-CB1 (at approximately 128 kDa) and anti-CB2-immunoreactive bands (at 38 and 26 kDa), which were abolished by preadsorption with the respective blocking peptide (lower lanes). Anti-ERK2 was used as an equal loading control. (O) In humans, pancreatic concentrations of the endocannabinoid anandamide (AEA) are higher in acute pancreatitis (black bars) than in normal pancreas (white bars; P < .05), whereas levels of combined 1- plus 2-arachidonoylglycerol (1 - AG + 2 - AG) are unchanged.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Immunohistochemical staining, Derivative Assay, Control, Immunostaining, Staining, Over Expression, Western Blot, Blocking Assay

Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Levels of serological, molecular, and inflammatory markers in acute pancreatitis and effects of CB1/CB2 antagonists. (A) Levels of serum lipase, pancreatic MPO, and the pancreatic wet weight/dry weight ratio are increased in mice following cerulein-induced pancreatitis (black bars) over control animals (white bars; * P < .05). (B) Pancreatic levels of IL-6 protein and KC1 mRNA (normalized to GAPDH expression levels) rise significantly following induction of acute pancreatitis. Treatment with cannabinoid receptor antagonists AM251 or AM630 either alone or in combination did not change levels of these parameters, except pancreatic KC1 mRNA expression (P < .05). * and # represent P < .05 as compared with the saline group or the cerulein group, respectively. (*) Mann–Whitney U test. (#) Analysis of variance followed by post hoc Bonferroni’s multiple comparison test. n.s. = statistically not significant.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Control, Expressing, Saline, MANN-WHITNEY, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: Effect of CB2, PPARα and PPARγ receptors antagonists on triglyceride accumulation per cell. Cells were incubated with different concentrations of BCP and 0.5 mM FFAm, in the presence or absence of specific receptor antagonists for 24 h. ( A ) Treatment of HepG2 cells with 5 µM CB2 antagonist AM630. ( B ) Treatment with 100 nM PPARα antagonist GW6471. ( C ) Treatment with 10 µM PPARγ antagonist GW9662. Data are expressed as a percentage change with respect to 0.5 mM FFA control condition (set equal to 100) and represent the mean ± SEM of five independent experiments. ** p < 0.01; *** p < 0.001 vs. BCP + 0.5 mM FFAm treated cells.

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Incubation, Control

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: qRT-PCR analysis of CB2 ( A ), PPARα ( B ) and PPARγ ( C ) mRNAs normalized for the housekeeping gene β-actin. Data are represented as the mean ± SEM of three independent experiments. * p < 0.05, *** p < 0.001 vs. control (untreated cells, set equal to 1) or vs. 0.5 mM FFAm-treated cells.

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Quantitative RT-PCR, Control

Journal: International Journal of Molecular Sciences

Article Title: Beta-Caryophyllene Modifies Intracellular Lipid Composition in a Cell Model of Hepatic Steatosis by Acting through CB2 and PPAR Receptors

doi: 10.3390/ijms24076060

Figure Lengend Snippet: Localization of CB2 receptors. Representative confocal images showing CB2 immunostaining (magenta) in HepG2 cell line. Nuclei are stained with DAPI (blue). Pictures are shown as max z-projections (low magnification; left ) with white arrows and arrowheads to highlight CB2high+ cells and CB2low+ cells, respectively, and a single confocal plane with reslicing ( right ) to better appreciate the intracellular distribution of CB2+ puncta in two of the cells present in the image. The cyan contoured image shows a cell positive for CB2 at low levels; the yellow contoured image identifies a cell with extensive immunolabelling for CB2. Scale bars: 50 µm (low magnification) and 10 µm (high magnification).

Article Snippet: Chemicals used were: (E)-β-caryophyllene (BCP) purchased from Sigma-Aldrich (St. Louis, MO, USA), AdipoRedTM assay reagent from Lonza (Walkersville, MD, USA), NucBlue Live ReadyProbes Reagent from Invitrogen (Carlsbad, CA, USA), CellTiter-Glo ® Luminescent Cell Viability from Promega (Madison, WI, USA), anti-CB2 primary antibody from Cayman chemical (Ann Arbor, MI, USA), anti-Rabbit IgG AlexaFluor647 secondary antibody from Jackson Immunoresearch (Ely, UK), AM630, GW9662 and GW6741 antagonists from Cayman chemical and sodium oleate, sodium palmitate and bovine serum albumin from Sigma-Aldrich.

Techniques: Immunostaining, 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: 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

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: Translational Lung Cancer Research

Article Title: Cannabinoid receptor 2 expression in early-stage non-small cell lung cancers identifies patients with good prognosis and longer survival

doi: 10.21037/tlcr-22-247

Figure Lengend Snippet: Patients’ characteristics categorized by gender, clinical stage, grading, and histology

Article Snippet: Samples were stained using mouse monoclonal anti-CB1 antibody (ImmunoGenes, Cat# 01, RRID:AB_2910137) and mouse monoclonal anti-CB2 antibody (Abnova, Cat# H00001269-M01, RRID:AB_875479) according to the manufacturer’s protocol.

Techniques: Paraffin-embedded Immunohistochemistry

Journal: Translational Lung Cancer Research

Article Title: Cannabinoid receptor 2 expression in early-stage non-small cell lung cancers identifies patients with good prognosis and longer survival

doi: 10.21037/tlcr-22-247

Figure Lengend Snippet: Kaplan-Meier analysis of OS, CSS, and DFS for CB2 gene (A) and protein (B) expression positivity in tumor tissue. CB2, cannabinoid receptor 2; qRT-PCR, quantitative reverse-transcriptase polymerase chain reaction; neg., negative; pos., positive; HR, hazard ratio; IHC, immunohistochemistry; OS, overall survival; CSS, cancer specific survival; DFS, disease free survival.

Article Snippet: Samples were stained using mouse monoclonal anti-CB1 antibody (ImmunoGenes, Cat# 01, RRID:AB_2910137) and mouse monoclonal anti-CB2 antibody (Abnova, Cat# H00001269-M01, RRID:AB_875479) according to the manufacturer’s protocol.

Techniques: Expressing, Quantitative RT-PCR, Reverse Transcription, Polymerase Chain Reaction, Immunohistochemistry

Journal: Translational Lung Cancer Research

Article Title: Cannabinoid receptor 2 expression in early-stage non-small cell lung cancers identifies patients with good prognosis and longer survival

doi: 10.21037/tlcr-22-247

Figure Lengend Snippet: Patients’ characteristics categorized by weight and BMI

Article Snippet: Samples were stained using mouse monoclonal anti-CB1 antibody (ImmunoGenes, Cat# 01, RRID:AB_2910137) and mouse monoclonal anti-CB2 antibody (Abnova, Cat# H00001269-M01, RRID:AB_875479) according to the manufacturer’s protocol.

Techniques: Paraffin-embedded Immunohistochemistry

Journal: Translational Lung Cancer Research

Article Title: Cannabinoid receptor 2 expression in early-stage non-small cell lung cancers identifies patients with good prognosis and longer survival

doi: 10.21037/tlcr-22-247

Figure Lengend Snippet: CB1 (A) and CB2 (B) gene expression (ΔCT values) in tumor and tumor-free lung tissue from 10 NSCLC patients. NSCLC, non-small cell lung cancer; CB1, cannabinoid receptor 1; CT, cycle threshold; CB2, cannabinoid receptor 2.

Article Snippet: Samples were stained using mouse monoclonal anti-CB1 antibody (ImmunoGenes, Cat# 01, RRID:AB_2910137) and mouse monoclonal anti-CB2 antibody (Abnova, Cat# H00001269-M01, RRID:AB_875479) according to the manufacturer’s protocol.

Techniques: Expressing

Journal: Translational Lung Cancer Research

Article Title: Cannabinoid receptor 2 expression in early-stage non-small cell lung cancers identifies patients with good prognosis and longer survival

doi: 10.21037/tlcr-22-247

Figure Lengend Snippet: Immunohistochemical staining of FFPE tumor tissue samples of NSCLC patients using anti-CB1 and anti-CB2 monoclonal antibodies (under ×10 lens). IHC, immunohistochemistry; CB1, cannabinoid receptor 1; CB2, cannabinoid receptor 2; FFPE, formalin-fixed paraffin-embedded; NSCLC, non-small cell lung cancer.

Article Snippet: Samples were stained using mouse monoclonal anti-CB1 antibody (ImmunoGenes, Cat# 01, RRID:AB_2910137) and mouse monoclonal anti-CB2 antibody (Abnova, Cat# H00001269-M01, RRID:AB_875479) according to the manufacturer’s protocol.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry, Formalin-fixed Paraffin-Embedded