Journal: Cells

Article Title: Adenosine A 2B Receptor Antagonism Interferes with TGF-β Cellular Signaling Through SMAD2/-3 and p65-Nf-κB in Podocytes and Protects from Phenotypical Transformation in Experimental Diabetic Glomerulopathy.

doi: 10.3390/cells14120890

Figure Lengend Snippet: Figure 2. Immunohistochemical analysis of glomerular alterations in diabetic rats. (A) The panels show representative immunofluorescent images of the detection of A2BAR, α-SMA, ZO-1, nephrin, and Snail in renal tissue from control rats, diabetic nephropathy (DN) rats, and diabetic rats treated with the A2BAR antagonist MRS1754 (DN + MRS1754). The images emphasize changes in the levels of these proteins within the glomerulus. Nuclear staining with DAPI is in blue. The bar indicates 50 µm. (B) The graphs correspond to the means ± SD of relative fluorescent units (RFUs) of a quantitative analysis of A2BAR, α-SMA, ZO-1, nephrin, and Snail in glomeruli from the control (white bar), diabetic (black bar), and MRS1754-treated diabetic (grey bar) rat groups. The means in the control group were normalized to 1. Images of 12 glomeruli from 2 control, 4 diabetic, and 4 treated diabetic rats were analyzed. *, p < 0.05 vs. control; #, p < 0.05 vs. diabetes.

Article Snippet: The samples were incubated with an antibody against A2BAR (dilution of 1:250, cat. orb234999, Biorbyt Ltd., Cambridge, UK), anti-α-SMA (dilution of 1:100, cat. sc-130617, Santa Cruz Biotech, Dallas, TX, USA), anti-ZO1 (dilution of 1:100, cat. 61-7300, Thermo Fisher Scientific, USA), anti-nephrin (dilution of 1:100, cat. PA5-106921, Thermo Fisher Scientific, USA), and anti-Snail (dilution of 1:100, cat. ab53519, Abcam, Cambridge, UK) overnight at 4 ◦C.

Techniques: Immunohistochemical staining, Control, Staining

Journal: Cells

Article Title: Adenosine A 2B Receptor Antagonism Interferes with TGF-β Cellular Signaling Through SMAD2/-3 and p65-Nf-κB in Podocytes and Protects from Phenotypical Transformation in Experimental Diabetic Glomerulopathy.

doi: 10.3390/cells14120890

Figure Lengend Snippet: Figure 3. A2BAR antagonism prevents the loss of podocyte epithelial markers in diabetic glomeru- lopathy. (A) Representative Western blot analysis showing the levels of epithelial ZO-1 and nephrin, as well as mesenchymal α-SMA and Snail, in purified glomeruli from the different rat groups. (B) The graphs represent the means and SD of the levels of epithelial and mesenchymal markers relative to β-actin. The means in the control group were normalized to 1. (C) Representative Western blot analysis of the Snail distribution in cell fractions purified from the kidneys of experimental rat groups. (D) The graph represents the ratio of the Snail subcellular distribution. The mean in the control was normalized to 1. The number of animals analyzed in each experimental group was 5 in (B) and 2 in (D). *, p < 0.05 vs. control; #, p < 0.05 vs. DN; &, p < 0.05 vs. control.

Article Snippet: The samples were incubated with an antibody against A2BAR (dilution of 1:250, cat. orb234999, Biorbyt Ltd., Cambridge, UK), anti-α-SMA (dilution of 1:100, cat. sc-130617, Santa Cruz Biotech, Dallas, TX, USA), anti-ZO1 (dilution of 1:100, cat. 61-7300, Thermo Fisher Scientific, USA), anti-nephrin (dilution of 1:100, cat. PA5-106921, Thermo Fisher Scientific, USA), and anti-Snail (dilution of 1:100, cat. ab53519, Abcam, Cambridge, UK) overnight at 4 ◦C.

Techniques: Western Blot, Purification, Control

Journal: Livers

Article Title: Translocation of Adenosine A2B Receptor to Mitochondria Influences Cytochrome P450 2E1 Activity after Acetaminophen Overdose

doi: 10.3390/livers4010002

Figure Lengend Snippet: Figure 4. A2BAR and PGRMC1 in the mitochondrial outer membrane. Mice were administered 300 mg/kg of APAP followed by BAY 60-6583 (4 mg/kg) or PSB 603 (1 mg/kg) 6 h later, and liver

Article Snippet: The following primary antibodies were used: A2BAR (#ab229671) and Cyp2E1 (#ab28146) from Abcam; VDAC (#4866), β-actin (#4970), -2 (#89326S) and PGRMC1 (#13856S) from Cell Signaling Technology; Flotillin (Sigma Aldrich #F1180) and Tom 20 (Santa Cruz Biotechnology sc-11415).

Techniques: Membrane

Journal: Livers

Article Title: Translocation of Adenosine A2B Receptor to Mitochondria Influences Cytochrome P450 2E1 Activity after Acetaminophen Overdose

doi: 10.3390/livers4010002

Figure Lengend Snippet: Figure 5. Putative A2BAR interaction with PGRMC1 and its influence on Cyp2E1 activity. (A) Surface representation of docked complex showing the interaction of A2BAR (green) with PGRMC1 (cyan) with a cleft formed by residues. Mice were administered a dose of 300 mg/kg of APAP, followed by treatment with PSB 603, 6 h later. After 24 h, liver tissue was collected, and the mitochondrial fraction was isolated. (B) Mitochondrial Cyp2E1 activity and (C) mitochondrial Cyp2E1 protein levels after APAP overdose. Data represent means ± SE of n = 3–4 animals per group. * p < 0.05 (compared with control), # p < 0.05 (compared to APAP).

Article Snippet: The following primary antibodies were used: A2BAR (#ab229671) and Cyp2E1 (#ab28146) from Abcam; VDAC (#4866), β-actin (#4970), -2 (#89326S) and PGRMC1 (#13856S) from Cell Signaling Technology; Flotillin (Sigma Aldrich #F1180) and Tom 20 (Santa Cruz Biotechnology sc-11415).

Techniques: Activity Assay, Isolation, Control