Journal: Journal of Cellular Physiology

Article Title: Cannabinoid Receptors Modulate Physiological Remodelling of the Blood–Testis Barrier

doi: 10.1002/jcp.70109

Figure Lengend Snippet: Gene expression analysis of OCLN and markers related to its trafficking in WT and CB1 −/− (KO) testes. RTqPCR analysis of Ocln mRNA. Transcript amounts were reported as normalized fold expression (nfe) relatively to Rps18 gene (A). Western blot analysis of OCLN. Protein amounts were quantified by densitometry analysis, normalized against ERK1/2 signals and expressed in OD values (B). Immunofluorescence analysis of OCLN; white arrowheads represent OCLN localization (green) in testis. Nuclei were labelled with DAPI (blue). Scale bar: 5 µm (C). Quantitative analysis of OCLN green fluorescence on supra‐basal epithelium Regions of Interest (ROI); values are expressed as Sum of Intensity/area (SUM (I)/pixel 2 ), measured using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) (D). RTqPCR analysis of endocytic (Eea1), recycling (Rab13) and degradation (Rab9 and Itch) markers of OCLN. Transcript amounts were reported as normalized fold expression (nfe) relatively to Rps18 gene (E). All data were reported as mean value ± S.E.M. (* p < 0.05; ** p < 0.01).

Article Snippet: The green fluorescence signal diffused in the luminal compartment was quantified using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) (see Figure ).

Techniques: Gene Expression, Expressing, Western Blot, Immunofluorescence, Fluorescence, Imaging, Software

Journal: Journal of Cellular Physiology

Article Title: Cannabinoid Receptors Modulate Physiological Remodelling of the Blood–Testis Barrier

doi: 10.1002/jcp.70109

Figure Lengend Snippet: Responsiveness of OCLN expression to CB1 and CB2 receptors. Western blot analysis of OCLN in testis of HT mice in vitro treated with vehicle (CTRL), ACEA (A) or JWH‐133 (B) at different concentrations: 0.1–1–10 µM. Protein amounts were quantified by densitometry analysis, normalized against ERK1/2 signals and expressed in OD as fold change. Immunofluorescence analysis of OCLN in testis of CTRL and ACEA (10 µM) or JWH‐133 (1 µM); white arrowheads represent OCLN localization (green) in testis. Nuclei were labelled with DAPI (blue). Scale bar: 5 µm (C). Quantitative analysis of OCLN green fluorescence on supra‐basal epithelium Regions of Interest (ROI); values are expressed as Sum of Intensity/area (SUM (I)/pixel 2 ), measured using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) (D). RTqPCR analysis of Ocln mRNA in testis of CTRL and ACEA 10 µM (E) or JWH‐133 1 µM (F). Transcript amounts were reported as normalized fold expression (nfe) relatively to Rps18 gene. All the data were reported as mean value ± S.E.M. (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; a vs. b: p < 0.05).

Article Snippet: The green fluorescence signal diffused in the luminal compartment was quantified using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) (see Figure ).

Techniques: Expressing, Western Blot, In Vitro, Immunofluorescence, Fluorescence, Imaging, Software

Journal: Journal of Cellular Physiology

Article Title: Cannabinoid Receptors Modulate Physiological Remodelling of the Blood–Testis Barrier

doi: 10.1002/jcp.70109

Figure Lengend Snippet: CB1 and CB2 action on BTB permeability and functionality. FITC‐dextran diffusion assay in testis of WT and HT mice in vitro treated with vehicle (CTRL), ACEA 10 µM and JWH‐133 1 µM (A) Scale bar: 300 μm. The green fluorescence signal diffused in the luminal compartment was quantified using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) as SUM (I)/pixel 2 (see also Figure for technical details) (B) a versus b: p < 0.05. RTqPCR analysis of endocytic (Eea1), recycling (Rab13) and degradation (Rab9 and Itch) markers of OCLN in the testis of CTRL and ACEA 10 µM (C) or JWH‐133 1 µM (D). Transcript amounts were reported as normalized fold expression (nfe) relatively to Rps18 gene. All data were reported as mean value ± S.E.M. (* p < 0.05; ** p < 0.01).

Article Snippet: The green fluorescence signal diffused in the luminal compartment was quantified using the Nikon Imaging Analytical Software (NIS‐Elements Analysis D 4.40.00, 64 bit) (see Figure ).

Techniques: Permeability, Diffusion-based Assay, In Vitro, Fluorescence, Imaging, Software, Expressing