Journal: bioRxiv

Article Title: Tandem Requirement for Full Renal D 1 R and D 5 R Activity

doi: 10.1101/736611

Figure Lengend Snippet: A: In the human renal cortex, D 1 R immunostaining was found throughout the cytoplasm, luminal, and basolateral membranes in renal proximal tubules (PT) and collecting tubule (CT). By contrast, C: D5R immunostaining was noted in luminal but not in basolateral membranes of renal proximal tubules (RPT) or collecting tubule (CT). E: D5R immunostaining was also noted in the medullary thick ascending limbs of Henle (red arrows) but not in medullary collecting ducts (blue arrows) or capillaries (green arrows). No staining was evident in slices with the D 1 R ( B ) and D 5 R ( D, F ) antibody preadsorbed with their respective immunizing peptides. D5R immunostaining was also noted in some of the cultured human renal proximal tubule cells (hRPTCs) (black arrows) ( G ). No staining was evident in cultured hRPTCs with the D 5 R antibody pre-adsorbed with the D 5 R immunizing peptide ( H ). Total cell lysates from hRPTCs treated with vehicle (0) or fenoldopam (1 μM/15 min) were immunoprecipitated (IP) with anti-D 1 R or anti-D 5 R antibodies and immunoblotted for D 5 R or D 1 R, respectively ( I, J ). Normal rabbit (Rb) IgG was used as immunoprecipitant for the negative control. A D 5 R-enriched HEK-293 cell lysate (Novus Biologicals Cat. #NBL1- 10019) (I) or a hRPTC lysate was used for regular immunoblot (IB). ( J )

Article Snippet: Since the D 1 -like receptors share almost the same anatomical distribution at the proximal tubules, we next determined whether the receptors interact in hRPTCs and observed that these receptors co-immunoprecipitated at the basal state (mostly as dimers) and upon treatment (both as monomers and dimers) with the D 1 -like (D 1 R/D 5 R) receptor agonist fenoldopam in D 5 R-enriched HEK293 cells (Novus Biologicals Cat. #NBL1-10019) and hRPTCs ( ).

Techniques: Immunostaining, Staining, Cell Culture, Immunoprecipitation, Negative Control, Western Blot

Journal: Molecular cell

Article Title: Dopamine Uses the DRD5-ARRB2-PP2A Signaling Axis to Block the TRAF6-Mediated NF-κB Pathway and Suppress Systemic Inflammation.

doi: 10.1016/j.molcel.2020.01.022

Figure Lengend Snippet: Figure 6. Inhibition of TLR2-Induced Inflammation by DRD5 Activation Requires the ARRB2/PP2A Signaling Complex (A) Left: qRT-PCR analysis of Il6 and Tnf mRNA expression in WT and Arrb2/ BMDMs treated as indicated for 6 h. ELISA of IL-6 and TNF-a in su- pernatants from WT and Arrb2/ BMDMs treated as indicated for 18 h. (B) Immunoblot analysis of p-IkBa, p-p38, and ARRB2 in lysates of WT and Arrb2/ BMDMs treated as indicated. (C) Left: qRT-PCR analysis of Il6 and Tnf mRNA expression in PP2Acf/f and PP2Acf/fLysM-Cre BMDMs treated as indicated for 6 h. Right: ELISA of IL-6 and TNF-a in supernatants from PP2Acf/f

Article Snippet: Gang Pei, Tongji University,China N/A Mouse:PP2Acfl/fl Dr. Xiang Gao, Nanjing University,China & Dr. Jun Gao, Nanjing Medical University,China N/A Mouse:LysM-Cre The Jackson Laboratory Stock No:004781 Mouse:Drd5 / Cyagen Biosciences Inc N/A Oligonucleotides See Table S1 for oligonucleotide information This paper N/A Recombinant DNA Plasmid: human DRD1 Dr. Jiawei Zhou, SIBS,China N/A Plasmid: human DRD2 Dr. Jiawei Zhou, SIBS,China N/A Plasmid: human DRD3 Addgene Cat#24098 Plasmid: human DRD4 Addgene Cat#24100 Plasmid: human DRD5 Addgene Cat#66272 Plasmid: mouse DRD1 OriGene Cat# MR226226 Plasmid: mouse DRD3 OriGene Cat# MR226047 Plasmid: mouse DRD5 OriGene Cat# MR222905 Plasmid: mouse ARRB2 OriGene Cat# MR206444 Plasmid: mouse PP2Ac Dr. Dong Xie,SIBS, China N/A Plasmid: human ARRB2 Dr.

Techniques: Inhibition, Activation Assay, Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Molecular cell

Article Title: Dopamine Uses the DRD5-ARRB2-PP2A Signaling Axis to Block the TRAF6-Mediated NF-κB Pathway and Suppress Systemic Inflammation.

doi: 10.1016/j.molcel.2020.01.022

Figure Lengend Snippet: Figure 7. DRD5-ARRB2-PP2A Signaling Acti- vation Prevents S. aureus-Induced Sepsis and Meningitis (A) Survival of WT, Drd5/, Arrb2/, and PP2Acf/f

Article Snippet: Gang Pei, Tongji University,China N/A Mouse:PP2Acfl/fl Dr. Xiang Gao, Nanjing University,China & Dr. Jun Gao, Nanjing Medical University,China N/A Mouse:LysM-Cre The Jackson Laboratory Stock No:004781 Mouse:Drd5 / Cyagen Biosciences Inc N/A Oligonucleotides See Table S1 for oligonucleotide information This paper N/A Recombinant DNA Plasmid: human DRD1 Dr. Jiawei Zhou, SIBS,China N/A Plasmid: human DRD2 Dr. Jiawei Zhou, SIBS,China N/A Plasmid: human DRD3 Addgene Cat#24098 Plasmid: human DRD4 Addgene Cat#24100 Plasmid: human DRD5 Addgene Cat#66272 Plasmid: mouse DRD1 OriGene Cat# MR226226 Plasmid: mouse DRD3 OriGene Cat# MR226047 Plasmid: mouse DRD5 OriGene Cat# MR222905 Plasmid: mouse ARRB2 OriGene Cat# MR206444 Plasmid: mouse PP2Ac Dr. Dong Xie,SIBS, China N/A Plasmid: human ARRB2 Dr.

Techniques: