Journal: Journal of hematology & oncology

Article Title: DT2216-a Bcl-xL-specific degrader is highly active against Bcl-xL-dependent T cell lymphomas.

doi: 10.1186/s13045-020-00928-9

Figure Lengend Snippet: Fig. 1 DT2216 selectively kills Bcl-xL-dependent TCL cell lines by degrading Bcl-xL in a VHL-dependent manner. a A represent immunoblot analysis of expression of various Bcl-2 family proteins and VHL in selective TCL cell lines. b Effects of DT2216 and ABT263 on MyLa cell and platelet (PLT) viability after 72 h treatment. The data are mean ± SEM (n = 3 independent assays). c DT2216 induced Bcl-xL degradation in a dose-dependent manner. MyLa cells were pretreated with or without QVD for 4 h, and then treated with DT2216 for 16 h. d DT2216 induced Bcl-xL degradation in a time-dependent manner. e ABT263 and/or the VHL ligand (VHL-L) could not induce Bcl-xL degradation. f ABT263 pretreatment blocked the degradation of Bcl-xL by DT2216. g VHL-L pretreatment blocked the degradation of Bcl-xL by DT2216. h MG132 blocked the degradation of Bcl-xL by DT2216. i DT2216 could not induce Bcl-xL degradation in VHL-null 786-O cells. j DT2216, but not its negative control compound (DT2216 NC), induced Bcl-xL degradation and cleavage of caspase-3 and PARP. For Fig. 1e–h, MyLa cells were pretreated with indicated compounds for 2 h, and proteins were measured 16 h after treatment with DT2216. k Pretreatment with VHL-L blocked the effect of DT2216 on cell viability. MyLa cells were pretreated with VHL-L for 2 h, and then treated with DT2216 for 72 h. The data are mean ± SD (n = 6 replicates). a p < 0.05 vs. DT2216. l DT2216, but not DT2216 NC, reduced the cell viability that was assayed after treatment with DT2216 or DT2216 NC for 72 h. The data are mean ± SD (n = 6 replicates). m DT2216-induced Bcl-xL degradation persisted up to 48 h upon the removal of DT2216 after 24 h DT2216 treatment. n The effect of DT2216, but not ABT263, on reducing MyLa cell viability, persisted up to 48 h upon the removal of ABT263 or DT2216 after 24 h treatment. The data presented are mean ± SD (n = 3 replicates)

Article Snippet: The primary antibodies including Bcl-xL (Cat. No. 2762), Mcl-1 (Cat. No. 5453), Bcl-2 (Cat. No. 4223), Bax (Cat. No. 2772), Bak (Cat. No. 12105), Bim (Cat. No. 2933), Noxa (Cat. No. 14766), VHL (Cat. No. 68547), Caspase-3 (Cat. No. 9662), cleaved Caspase-3 (Cat. No. 9661), PARP (Cat. No. 9532), β-actin (Cat. No. 4970), and the secondary horse radish peroxidase (HRP)-linked antibody (Cat. No. 7074) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Western Blot, Expressing, Negative Control

Journal: Cell stem cell

Article Title: Vimentin coordinates protein turnover at the aggresome during neural stem cell quiescence exit

doi: 10.1016/j.stem.2020.01.018

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Harm Kampinqa N/A Tdp43-gfp overexpression Addgene Cat#28197 mNeon overexpression Gift from Dr. Erik Dent, mNeon cDNA from Allele Biotechnology & Pharmaceuticals, Inc N/A VHL-BFP overexpression This paper, VHL insert from Addgene #21053 N/A GFP-Ubiquitin Addgene Cat#11928 mCherry-GFP-LC3 Gift from Dr. Ashley Webb N/A psPAX2 Addgene Cat#12260 pCMV-VSV-G Addgene Cat#8454 Software and Algorithms Microsoft Excel Microsoft N/A Prism GraphPad N/A Adobe Illustrator Adobe.com N/A Imaris Oxford Instruments N/A Other N/A N/A N/A Open in a separate window KEY RESOURCES TABLE EXPERIMENTAL MODEL AND SUBJECT DETAILS Mice In this study, male and female mice between the ages of 6 weeks and 9 months were used.

Techniques: Immunofluorescence, Western Blot, Proximity Ligation Assay, Recombinant, SYBR Green Assay, RNA Sequencing Assay, Clone Assay, Over Expression, Software