Research Paper Volume 11, Issue 23 pp 11136—11147

Figure 2. Ninj2 shRNA or KO inhibits human glioma cell survival. A172 glioma cells (A–F), U251MG glioma cells (G and H) or the primary human glioma cells (derived two patients, “P1/P2”, G and H) 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 genes was tested by qPCR and Western blotting (A–C); Cell survival (D and G), soft agar colony formation (E) and cell death (F and H) were tested by appropriate assays. Ninj2 and Ninj1 proteins were quantified and normalized to the loading control (C). For each assay, n=5. For all the functional assays exact same number of viable cells of different genetic manipulations were initially seeded into each well/dish (At 0h or Day0) (same for all Figures). “Pare” stands for the parental control cells (same for all Figures). *p<0.05 vs. “Pare” cells. Experiments in this figure were repeated four times, and similar results were obtained.