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. 2016 Nov 29;7(52):87431–87448. doi: 10.18632/oncotarget.13704

Figure 5. Knockout of Sema3E in a PDAC cell line decreases cell growth and proliferation, as well as cell migration.

Figure 5

Sema3E-knockout PDAC cells were generated with Panc-28 cells using the CRISPR-Cas9 system, with sgRNA targeting SEMA3E gene. A. Immunoblot analysis shows the absence of the 87 kDa full-length protein in Panc28 Sema3E-knockout (Panc28-S3EKO) cells, compared to the parental wild type cells (Panc28-WT), confirming successful knockout of Sema3E protein. B. MTT assay shows that Panc28-S3EKO cells had significantly reduced cell growth over the period of 6 days compared to the WT control (two-way ANOVA and Bonferroni posttest, ***p<0.001). C. Cell cycle analysis shows a decreased rate in cell proliferation in Panc28-S3EKO cells compared to the control, as indicated by a significant S to G1 phase shift (Student's t-test, **p<0.01). D. A wound healing scratch assay shows that the Sema3E-knockout cells had less cell movement towards the center of the wound scratch, compared to the parental wild type control (two-way ANOVA and Bonferroni posttest,*p<0.05). Red squares are small sections in the wound area that got further magnified to demonstrate less wound closure in Panc28-S3EKO cells vs. the control cells Panc28-WT. All data are represented as mean ± S.E.M, and are representative of at least 3 independent experiments.