Fig. 5

KPNB1 interacted with YBX1 and regulated its nuclear localization in glioblastoma. A. Mass spectrometry was used to identify the proteins obtained in U251MG cells. A subset of the identified peptides is displayed. B. Mass spectrometric sequencing of YBX1. C. Western blot analysis of reciprocal co-IP in U87MG and U251MG cells, demonstrating that endogenously expressed KPNB1 bound to YBX1. D. KPNB1-Flag expression plasmid was transfected into 293 T cells. Reciprocal co-IP demonstrating the interaction of exogenously expressed KPNB1 with endogenously expressed YBX1. E-F. Western blot analysis was used to detect the protein expression of YBX1 in the cytoplasm and nucleus of U87MG and U251MG infected with sh-Control or shKPNB1 (E) and EV or KPNB1 plasmids (F). G. U251MG cells were classified as treatment (shKPNB1) or control. Cells were incubated with rabbit anti-KPNB1 antibody and mouse anti-YBX1 antibody for immunofluorescence to determine the nuclear translocation of YBX1. H. PLA was used to verify the interaction between KPNB1 and YBX1 in U251MG cells. The size of the scale bar in microscopy images was 20 μm