Figure 4. Ectopic overexpression of NINJ2 promotes CRC cell survival and proliferation. The lentivirus encoding NINJ2 cDNA construct was added to HT-29 cells. Via selection by puromycin two stable cell lines (“Line-1/-2”) were established. Control cells were infected with virus encoding empty vector (“Vec”); Expression of NINJ2 mRNA (A), NINJ1 mRNA (B) and listed proteins (C) were shown; Cells were further cultured for the indicated time periods, cell survival was tested by MTT assay (D); Cell proliferation was tested by BrdU incorporation assay (E) and soft agar colony formation assay (F); The primary human colon cancer cells (“pri-Can-1/-2/-3”) were infected with lentivirus encoding NINJ2 cDNA construct (“NINJ2-OE”) or the empty vector (“Vec”); Cells were cultured for the indicated time periods; NINJ2 mRNA expression was tested by qPCR assay (G); Cell survival and proliferation were tested by the MTT assay (H) and BrdU incorporation assay (I), respectively. For each assay, n=5. NINJ1 and NINJ2were normalized to the loading control Tubulin (C). * P< 0.05 vs.“Vec” cells. Experiments in this figure were repeated three times, and similar results were obtained.