SRPIN340
National Institutes of Health
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Alternative splicing is a fundamental phenomenon to build protein diversity in health and diseases. Extra Domain A+ Fibronectin…
.................................................................................................................................. viii RESUMO ........................................................................................................................................ ix Chapter 1: General Introduction .................................................................................................. 1 1. General Introduction ............................................................................................................. 2 1.1 Alternative splicing and its role in cancer ..................................................................... 2 1.2 SR Proteins and their roles in pre-mRNA splicing ......................................................... 4 1.3 SRPK and their oncogenic activity ................................................................................. 5 1.4 SRPK as drug target ....................................................................................................... 9 1.5 Aims of the thesis ........................................................................................................ 11 Chapter 2: Research Article I ...................................................................................................... 12 2. Research Article I: Potential Antileukemia Effect and Structural Analyses of SRPK Inhibition by N-(2-(Piperidin-1-yl)-5-(Trifluoromethyl)Phenyl)Isonicotinamide (SRPIN340) ................. 13 2.1 Abstract ....................................................................................................................... 14 2.2 Introduction ................................................................................................................ 15 2.3 Experimental procedures ............................................................................................ 16 2.3.1 Cell lines .............................................................................................................. 16 2.3.2 Isolation of PBMC from human blood ................................................................ 17 2.3.3 SRPK inhibitor synthesis ...................................................................................... 17 2.3.4 MTT cell viability assay ....................................................................................... 17 2.3.5 Flow cytometry assays ........................................................................................ 17 2.3.6 RT-PCR and RT-qPCR analysis ............................................................................. 17 2.3.7 Western blot analysis ......................................................................................... 18 2.3.8 Cloning, expression and purification procedures ............................................... 18 2.3.9 Fluorescence spectroscopy ................................................................................. 19 2.3.10 Computational analysis ..................................................................................... 19 2.3.11 Statistical analysis ............................................................................................. 20 2.4 Results ......................................................................................................................... 21 2.4.1 Expression of SRPK1 and SRPK2 in leukemia cells .............................................. 21
Purpose AMD is a major cause of blindness. Current therapy aims to suppress VEGF levels in order to prevent CNV, which causes AMD…
Purpose To investigate the applicability of serine/arginine-rich protein kinase (SRPK)-specific inhibitor, SRPIN340, for…