Research Paper Volume 11, Issue 23 pp 11136—11147

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.