Fig. 1: SAMHD1 interacts with NONO.
From: Stabilization of SAMHD1 by NONO is crucial for Ara-C resistance in AML
A, B Validation of the exogenous interaction between NONO and SAMHD1 by co-IP. HA-tagged SAMHD1 was cotransfected with Myc-tagged NONO into 293 T cells. Co-IP was performed with anti-HA or anti-Myc agarose, and the interactions were examined by western blotting. C, D Identification of the endogenous interaction between NONO and SAMHD1 in HL60 and THP-1 cells. Cell lysates were immunoprecipitated with an anti-NONO antibody, and the indicated proteins were examined by western blotting. IgG was used as the negative control. E NONO interacts with SAMHD1 detected by GST pulldown assay. E. coli-expressed GST-NONO, GST-SAMHD1, or GST (as a negative control) proteins were immobilized by Glutathione-Sepharose and then incubated with THP-1cell lysate. GST or GST-NONO, GST-SAMHD1 and pull downed proteins were examined by western blotting with indicated antibodies. F Identification of the co-location of NONO and SAMHD1 in THP-1 cells by immunofluorescence. GFP-SAMHD1 was cotransfected with Dsred-NONO into THP-1 cells for 24 h. Scale bar, 5 μM. G Schematic diagram of NONO and SAMHD1 truncation mutants. H The domain of SAMHD1 containing amino acids 115-562 interacts with NONO. HA-tagged NONO was cotransfected into 293 T cells with wild-type or mutant Myc-SAMHD1, and the interactions were verified by co-IP and western blotting. I The RRM domain of NONO interacts with SAMHD1. HA-tagged SAMHD1 was cotransfected into 293 T cells with wild-type or mutant Myc-NONO, and the interactions were verified by co-IP and western blotting.
