Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative Pinceaux staining showing a PC Soma (green, calbindin) along with the characteristic pinceaux formation on the axon side (red, CB1R) in both genotypes. B. Quantification of average CB1R pinceaux intensity across both genotypes and slide sets (VGAT, VGLUT1, VGLUT2). C. Same as B, but area of pinceaux. D-F. Proportion of CB1R puncta that colocalize with the respective vesicular marker before and after a 100 pixel shift of the CB1R channel along the X axis.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining, Marker

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative confocal images from WT, DMD mdx and CB1R KO cerebella demonstrating calbindin (green), raw CB1R (red), and filtered CB1R puncta (see methods: image analysis). The right column shows an expanded and merged view of the area indicated in the filtered CB1R column. Quantification of mean CB1R puncta intensity ( B ), density ( C ), and area ( D ) in WT (black) and DMDmdx (blue) cerebella.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques:

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGAT (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGAT image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGAT puncta in the molecular layer. E. Percentage of VGAT puncta colocalized with CB1R puncta. F. Intensity of CB1R puncta colocalized with VGAT puncta. G. Intensity of CB1R puncta not colocalized with VGAT puncta.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGLUT1 (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGLUT1 image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGLUT1 puncta in the molecular layer. E. Percentage of VGLUT1 puncta associated with CB1R puncta. F. Intensity of CB1R puncta colocalized with VGLUT1 puncta. G. Intensity of CB1R puncta not colocalized with VGLUT1 puncta

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: bioRxiv

Article Title: Loss of dystrophin reduces CB1 receptor expression and endocannabinoid-dependent synaptic plasticity in the cerebellar cortex

doi: 10.64898/2026.03.20.713279

Figure Lengend Snippet: A. Representative images demonstrating calbindin (green), filtered CB1R (red), and filtered VGLUT2 (cyan) staining across WT and DMD mdx cerebella. The right column shows an expanded and merged view of the area indicated in the VGLUT2 image. Quantification of mean intensity ( B ), density ( C ), and area ( D ) of VGLUT2 puncta in the molecular layer. E. Percentage of VGLUT2 puncta associated with CB1R puncta. F. Average intensity of CB1R puncta colocalized with VGLUT2 puncta. G. Intensity of CB1R puncta not colocalized with VGLUT2 puncta.

Article Snippet: Slides were incubated for 48 hours at 4°C under agitation with the following primary antibodies in blocking buffer: calbindin (1:3000, Sigma-Aldrich, Cat# C9848), CB1R (1:1000, Cayman Chemical Company, Cat# 10006590), and one of the following vesicular transporters - VGAT (1:250, Synaptic Systems, Cat# 131004), VGLUT2 (1:500, Synaptic Systems, Cat# 135418), or VGLUT1 (1:5000, Synaptic Systems, Cat# 135304).

Techniques: Staining

Journal: Molecular Medicine

Article Title: Progressive endocannabinoid system dysregulation in autosomal dominant polycystic kidney disease

doi: 10.1186/s10020-026-01457-w

Figure Lengend Snippet: ECS components are progressively dysregulated in human ADPKD kidney tissue. a Microarray analysis ( GSE7869 ) of human kidney tissue shows stepwise increases in CNR1 transcript from healthy cortex to minimally cystic (PKDm) and fully cystic (PKD) ADPKD tissue, with corresponding reductions in AEA-metabolizing enzymes NAPEPLD and FAAH . b Single-nucleus RNA-sequencing (snRNA-seq) analysis of human ADPKD kidneys ( n = 8) versus healthy controls ( n = 5) demonstrate consistent CNR1 upregulation and marked downregulation of NAPEPLD and FAAH , while 2-AG-metabolizing enzymes remain largely unchanged. c snRNA-seq analysis of diabetic kidney disease (DKD; n = 5 patients; controls n = 6) reveals minimal alterations in CNR1 and ECS-metabolizing enzymes. d Gene expression analysis by qPCR confirms CNR1 upregulation in human ADPKD kidney tissue ( n = 17) versus non-cystic nephrectomy controls ( n = 5), with concurrent changes in ECS enzyme transcription. e-i eCB quantification by liquid chromatography-tandem mass spectrometry (LC–MS/MS) reveals significant depletion of tissue anandamide (AEA); e , N -oleoylethanolamine (OEA); f , 2-arachidonoylglycerol (2-AG); h , and arachidonic acid (AA); i , while N -palmitoylethanolamine (PEA); g remains unchanged. j-l Western blot analysis shows substantial inter-individual variability in CB 1 R protein levels without significant difference between ADPKD ( n = 6) and control kidneys ( n = 4), but significant reductions in DAGLα/β, NAPEPLD, MGLL and FAAH protein. Western blots were normalized to total proteins. Data represent mean ± SEM. Statistics of control versus PKD represented by * and control versus PKDm by #. Statistical significance assessed by Mann–Whitney U test or unpaired t -test: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Sections were stained with rabbit anti-CB 1 R antibody (1:200, 10006590, Cayman) followed by a goat anti-rabbit HRP conjugate (ImmPRESSTM, Vector laboratories).

Techniques: Microarray, RNA Sequencing, Gene Expression, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Western Blot, Control, MANN-WHITNEY

Journal: Molecular Medicine

Article Title: Progressive endocannabinoid system dysregulation in autosomal dominant polycystic kidney disease

doi: 10.1186/s10020-026-01457-w

Figure Lengend Snippet: Temporal endocannabinoid system dysregulation during Pkd1 RC/RC disease progression. a-g Quantitative PCR analysis of ECS-related genes across disease stages (9 and 12 months) showing progressive upregulation of Cnr1 a and Cnr2 b transcripts, with corresponding changes in ECS metabolic enzymes: Dagla c , Daglb d , Napepld e , Mgll f , and Faah g . h-n Western blot analysis and quantification of ECS proteins across disease stages, demonstrating sustained CB 1 R protein elevation at 9 and 12 months h, i , stable 2-AG-related enzyme proteins (DAGLα, DAGLβ, MGLL; h, j, k, m ), and stage-specific changes in AEA-related enzymes including elevated NAPEPLD at 12 months ( h, l ) and reduced FAAH at 9 months ( h, n ). Data represent mean ± SEM from WT ( n = 5) and Pkd1 RC/RC ( n = 6–7) per group per timepoint. Statistical analysis: unpaired t -test comparing Pkd1 RC/RC to age-matched WT. Statistical significance: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus WT

Article Snippet: Sections were stained with rabbit anti-CB 1 R antibody (1:200, 10006590, Cayman) followed by a goat anti-rabbit HRP conjugate (ImmPRESSTM, Vector laboratories).

Techniques: Biomarker Discovery, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cancer Research Communications

Article Title: ROME, an Ancient Gene with a Novel Function in Vertebrates, Is a Key Modulator of Embryonal Development and Cancer Metastasis

doi: 10.1158/2767-9764.CRC-26-0068

Figure Lengend Snippet: ROME protein negatively regulates the Wnt pathway and calcium signaling in human cancer cells. A and B, Compared with EV control cells, TC-71 ( A ) and TC-32 ( B ) cells stably overexpressing ROME were less responsive to recombinant human Wnt3a protein as measured by TOPFlash β-catenin–responsive luciferase reporter. C, TC-71 cells with ROME knockdown by siRNA were more responsive to Wnt3a. D, Endogenous Wnt pathway activity in the colorectal cancer cell line HCT116 was significantly decreased by ROME expression (Western blots confirming ROME-HA expression on the right). Statistical significance was calculated by an unpaired t test (two-tailed) for A–D . E, ROME co-immunoprecipitates with β-catenin in β-catenin pulldown experiments. F, β-Catenin co-immunoprecipitates with ROME in ROME pulldown experiments. G, SPR sensorgrams showing direct binding between the recombinant full-length ROME protein and β-catenin protein. β-Catenin was immobilized on the surface and full-length ROME protein was injected in duplicate at concentrations of 1,250, 416.7, 138.9, 46.3, 15.4, and 5.12 nmol/L. The red lines are the actual data, and the black lines indicate the curve fit. H, ROME expression decreases β-catenin and TCF4 interaction (measured by co-IP). I–K, The Ca 2+ response was decreased in A4573, STA-ET-7.2, and TC-71 cells overexpressing ROME compared with that in EV-transfected cells when the cells were stimulated with ATP or FBS after overnight serum starvation. L, The Ca 2+ response was greater in the TC-71 cells with ROME KO than in the WT cells. Statistical significance was calculated by an unpaired t test (two-tailed) for I–L .

Article Snippet: For experiments involving binding of ROME-FL to vimentin (Novus Biologicals, #NBP2-35139-100 μg) and β-catenin (Novus Biologicals, #NBP3-18198-100 μg), vimentin and β-catenin were immobilized as ligands onto CM5 chip surfaces.

Techniques: Control, Stable Transfection, Recombinant, Luciferase, Knockdown, Activity Assay, Expressing, Western Blot, Two Tailed Test, Binding Assay, Injection, Co-Immunoprecipitation Assay, Transfection