Fig. 5

HNRNPK binds with ILF2 to promote the stabilization of ILF2. A-B Coimmunoprecipitation (Co-IP) assays were conducted using anti-HNRNPK (A) or anti-ILF2 (B) antibodies in TNBC cells, followed by immunoblot (IB) analysis for HNRNPK and ILF2. Immunoglobulin G (IgG) was utilized as a negative control antibody for immunoprecipitations. C-F Co-IP and IB assays demonstrated that LINC00571 overexpression led to an increase in the binding of HNRNPK and ILF2 (C-D), while LINC00571 knockdown resulted in decreased HNRNPK and ILF2 binding (E-F). G Immunoblot analysis depicted the cycloheximide (CHX) chase analysis of ILF2 protein degradation at indicated time points (t=0, 4, 8, 12h) in TNBC cells with or without HNRNPK. H Immunoblot analysis revealed the levels of ILF2 and HNRNPK in HNRNPK knockdown cells treated with vehicle control or MG132 (10 μM) for 12 hours in MDA-MB-231 cells (left) and BT-549 cells (right). I IP and IB demonstrated that knockdown of HNRNPK inhibited the ubiquitination of ILF2 in MDA-MB-231 cells (left) and BT-549 cells (right) treated with MG132. J IP and IB assays illustrated that overexpression of HNRNPK promoted the ubiquitination of ILF2 in MDA-MB-231 cells (left) and BT-549 cells (right) treated with MG132