Figure 6. Ninj2 associates with multiple receptor tyrosine kinases, required for downstream Akt and Erk activation. In A172 cells and primary human glioma cells (“P1/P2”) the association between Ninj2 with RTKs (EGFR, PDGFRβ and FGFR) was tested by co-immunoprecipitation (“Co-IP”) assays (A); “INPUT” shows expression of the RTKs and Ninj2 in total cell lysates (B). A172 cells were transduced with lentiviral Ninj2 shRNAs (“sh-Ninj2”, two different sequences “Seq1/Seq2”), control shRNA (“sh-C”) or the CRISPR/Cas9 Ninj2 KO construct (“ko-Ninj2”), stable cells were established via puromycin selection; expression of listed proteins was tested by Western blotting (C and D); Fresh human glioma tissue lysates from three patients (“T1/2/3”) were subjected to the same Co-IP assay (E), “INPUT” shows expression of RTKs and Ninj2 in the tumor lysates (F). Experiments in this figure were repeated five times, and similar results were obtained.