Research Paper Volume 11, Issue 19 pp 8526—8541

Figure 5. NINJ2 forms a complex with multiple receptor tyrosine kinases (RTKs) in CRC cells and colon cancer tissues. In HT-29 cells and primary human colon cancer cells (“pri-Can-1”) the associations between NINJ2 with multiple RTKs (EGFR, PDGFRα, PDGFRβ and FGFR) were tested by co-immunoprecipitation (Co-IP) assays (A); “Input” shows expression of RTKs and NINJ2 in total cell lysates (A). Fresh human colon tissue lysates from patient-1/-2/-3/ (“Pat-1/2/3”) were subjected to the same Co-IP assay of NINJ2-RTKs associations (B), “Input” shows expression of RTKs and NINJ2 in lysates (B). Expression of the listed proteins in stable HT-29 cells with applied NINJ2 shRNA (“Seq1/2/3”) or non-sense control shRNA (“shC”) were shown (C). Stable HT-29 cells with the lenti-CRISPR/Cas9-KO constructs (with NINJ2 sgRNA1/2, (D) or NINJ2 cDNA construct (“NINJ2-OE”, two lines, E) were subjected to the same Western blotting assay of listed proteins. NINJ2-bound RTKs (EGFR, PDGFRα, PDGFRβ and FGFR) were quantified (A and B). Akt and Erk phosphorylations were normalized to total proteins (C–E). Experiments in this figure were repeated three times, and similar results were obtained.