Journal: Nature

Article Title: Structural mechanism of angiogenin activation by the ribosome.

doi: 10.1038/s41586-024-07508-8

Figure Lengend Snippet: Fig. 1 | Angiogenin inhibits translation by binding to 80S ribosomes. a, Angiogenin (Ang) inhibits translation in RRL in a concentration-dependent manner. Inhibition is relieved by RNasin (RNase inhibitor). The apparent translation rate is shown (relative light units (RLU) s−1). Data are mean ± s.e.m. n = 2 or 3 independent experiments, as shown. b, Incubation of RRL with angiogenin for 15 min leads to the accumulation of tRNA fragments; fragment accumulation is inhibited by RNasin. n = 2 independent experiments. nt, nucleotides. c, Angiogenin co-pellets with the ribosome fraction from RRL, except in the presence of RNasin. n = 2 independent experiments. d, Cryo-EM analysis of RRL with 1 µM angiogenin reveals that over 30% of 80S ribosomes have a novel density in the A site. e, Rigid-body fit of human angiogenin (PDB: 5EOP)61 into the 3.4 Å cryo-EM map of angiogenin-bound ribosomes in RRL. f, A 3.0 Å cryo-EM map of in vitro assembled 80S–angiogenin complex. The 60S subunit is coloured cyan; 40S subunit is coloured yellow; P-site tRNA is coloured

Article Snippet: For anti-RNase A blotting, procedures were amended to block with 1× PBST with 0.1 mg ml−1 BSA and detected with 1:1,000 rabbit anti-bovine pancreatic RNase A polyclonal antibody-peroxidase conjugated (200-4388S, Rockland Immunochemicals).

Techniques: Binding Assay, Concentration Assay, Inhibition, Incubation, Cryo-EM Sample Prep, In Vitro

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: Total cellular RNA was prepared from EAhy926 and blood cells, and RT-PCR was performed as described in Materials and Methods. Densitometric analysis of the gels were used to represent the mRNA ratios by using Scion Image PC (National Institutes of Health). Results were normalized to the expression levels (E) of GAPDH and expressed as the ratio E (target)/E (GAPDH) (target = RNase 1 or RI). Each value represents the mean ± SD (n = 6) compared with EAhy926 cells. * indicates a significant differences with p<0.05, whereas ** indicates a significant difference with p<0.01.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: (A, B) Supernatants from the different cell types were prepared and analyzed by Western blot as described in Materials and Methods with antibodies against RNase 1 (poly) and RI (mono). Lane 1. positive control (A: 10 ng (0.56 pmol) of RNase 1, B: 30 ng (0.61 pmol) of RI), 2. EAhy926 cells, 3. platelets, 4. MNCs, 5. PMNs, 6. RBCs.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques: Western Blot, Positive Control

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: After fixation and blocking, cells were stained with antibodies against RNase 1 (green), RI (red), and nuclei (blue). Each image of platelets-2 and thrombin-activated platelets-2 is optical micrograph of platelets-1 and thrombin-activated platelets-1. Scale bars = 10 μm.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques: Blocking Assay, Staining

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: After fixation and blocking, cells were stained with antibodies against RNase 1 (green), VWF (red), and nuclei (blue). Scale bars = 10 μm.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques: Blocking Assay, Staining

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: EAhy926 cells (A) or platelets (C) are TEM images. EAhy926 cells (B) or platelets (D) immunostained with anti-RNase 1 (arrows) and anti-VWF antibodies (arrowheads). Scale bars (A) 2 μm, (B) 100 nm, (C) 500 nm, (D) 100 nm.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques:

Journal: PLoS ONE

Article Title: The expression and localization of RNase and RNase inhibitor in blood cells and vascular endothelial cells in homeostasis of the vascular system

doi: 10.1371/journal.pone.0174237

Figure Lengend Snippet: EAhy926 cells (A, B) or platelets (C, D) immunostained with anti-RNase 1 (arrows, (A, C)) or anti-VWF antibodies (arrowheads, (B, D)). Scale bars = 500 nm.

Article Snippet: Purified human RNase 1 (13468-H08H) and RI protein (M0307S) were from Sino Biologicals (Peking, PR China) and New England Biolabs Japan (Tokyo, Japan), respectively.

Techniques: