Journal: Cell reports

Article Title: Impact of WIN site inhibitor on the WDR5 interactome

doi: 10.1016/j.celrep.2020.108636

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: PolyScreen PVDF Hybridization Transfer Membrane , PerkinElmer , Cat# NEF1002.

Techniques: Western Blot, Recombinant, Protease Inhibitor, Staining, Clone Assay, Expressing, Transfection, Mass Spectrometry, Plasmid Preparation, Mutagenesis, CRISPR, Amplification, Modification, Software, Hybridization

Journal: Kidney360

Article Title: Mannose Binding Lectin Is Hydroxylated by Collagen Prolyl-4-hydroxylase and Inhibited by Some PHD Inhibitors

doi: 10.34067/KID.0000092020

Figure Lengend Snippet: Some PHD inhibitors inhibit MBL oligomerization. (A) Representative Western blot showing the effects of PHD inhibitors (roxadustat [Rox], molidustat [Mol], and vadadustat [Vad], all at 10 µM) on expression of MBL, P4HA1, HIF1α, and α-tubulin. (B) Secreted MBL in light fraction (F1–F5) or heavy fraction (F6–F10) separated by sucrose gradient after 24-hour treatment with respective PHD inhibitors. (C) Quantification of the percentage of secreted MBL in the heavy fraction (compared to total MBL) in samples treated with different PHD inhibitors is shown. **P<0.01, ***P<0.001. (D) Representative distribution of relative proportion of secreted MBL (normalized to total secreted MBL) in ten fractions separated by sucrose density gradient for one sample from each group. Uncropped blots are provided in Supplemental Figure 6.

Article Snippet: Dimethyloxaloylglycine (DMOG; catalog number A4506) and molidustat (catalog number BAY85-3934) were purchased from ApexBio Technology.

Techniques: Western Blot, Expressing

Journal: Kidney360

Article Title: Mannose Binding Lectin Is Hydroxylated by Collagen Prolyl-4-hydroxylase and Inhibited by Some PHD Inhibitors

doi: 10.34067/KID.0000092020

Figure Lengend Snippet: Prolyl hydroxylation in the collagen-like region is significantly inhibited by roxadustat and vadadustat but not molidustat. Cells were treated with 10 µM of roxadustat, vadadustat, and molidustat for 24 hours. (A) Representative Western blots showing the effects of 24 hours of treatment with different PHD inhibitors on various proteins of interest in the lysates of MBL-expressing HEK293S cells. (B) Representative Western blots showing the effects of 24 hours of treatment with different PHD inhibitors on MBL secretion in the supernatant. The supernatant from three biologic replicates was concentrated and mass spectrometry analysis on the samples without knowledge of the treatments was undertaken. (C) Relative area of prolyl hydroxylated peptides covering the collagen-like domain containing hydroxylation at various proline residues (normalized to total area of MBL peptides within the collagen-like domain) in different conditions is shown. (D) Relative area of lysyl-modified (open circles) and unmodified (open squares) peptides covering the collagen-like domain (normalized by total area of MBL peptides within the collagen-like domain) in different conditions is shown. **P<0.01, ***P<0.001. Uncropped blots are provided in the Supplemental Figure 8.

Article Snippet: Dimethyloxaloylglycine (DMOG; catalog number A4506) and molidustat (catalog number BAY85-3934) were purchased from ApexBio Technology.

Techniques: Western Blot, Expressing, Mass Spectrometry, Modification