Figure 3. NINJ2 knockout inhibits HT-29 cell survival and proliferation. HT-29 cells were transfected with the lenti-CRISPR/Cas9-KO constructs, with non-overlapping sgRNAs against NINJ2 (“sgRNA1/2”) or the non-sense control sgRNA (“sg-C”), stable cells were established via FACS sorting plus puromycin selection; Expression of NINJ2 mRNA (A), NINJ1 mRNA (B) and listed proteins (C) were shown; Cells were 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); Cell apoptosis was tested by the Annexin V-PI FACS assay (G) and TUNEL staining assay (H). NINJ1 and NINJ2were normalized to the loading control Tubulin (C). For each assay, n=5. * P< 0.05 vs. “sg-C” cells. Experiments in this figure were repeated three times, and similar results were obtained.