Fig 2: DANCR KO suppresses cell proliferation and stemness-like properties of pancreatic cancer cells.a DANCR KO suppresses cell proliferation in BXPC-3 and SW1990 cells, as determined by CCK8 assays. b DANCR KO suppresses colony formation. c DANCR KO inhibits sphere formation in number and size. d DANCR KO suppresses stemness-related genes in pancreatic cancer cells, as determined by qRT-PCR and at the protein level e. f Percentage of CD24+ and CD133+ population in DANCR KO and control (NC) cells, as determined by flow cytometry. The statistical results of flow cytometry were showed on the right. Values in a, b, d, e, f are mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Fig 3: Decreased Dab2 expression leads to altered expression of CSC markers. (a,b) Cell surface expression of CD133, CD24 and CD44 was determined by flow cytometry in COLO 357 and COLO357 1059 cells as described in the Methods section. Shown is a representative histogram depicting expression levels. Numerical values of expression are provided in Table 1. (c) mRNA expression for Bmi-1, Oct4, Nanog and Sox2 was determined by qRT-PCR analysis in untreated COLO357 Neg, COLO357 1059, Panc1 Neg and Panc1 1059 cells or cells treated with 2.5 ng/mL TGFß1 for 72 hrs. Relative expression is shown, with mRNA levels in the COLO357 Neg and Panc1 Neg cell lines for each gene set to 1.0 to allow comparison to their corresponding knockdown cell lines. Shown is the mean relative expression level ± SD performed in triplicate. * signifies p < 0.05 difference in expression for comparison to untreated Neg cell line; # represents p < 0.05 difference in expression for comparison to TGFß1-treated Neg cell line.

Fig 4: EMP3 interferes with stem-like properties.A CCLE database was interrogated for EMP3, SOX2, CD133, and EPCAM expression. Correlation between two genes in a total of 57 breast cancer cell lines was analyzed by Pearson statistics. B SOX2, CD133, and EPCAM expression in MDA-MB-231 cells were detected by real-time PCR. Mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001, vs. shCtrl. C Fractions of CD133-positive MDA-MB-231 cells were detected by flow cytometry. Mean ± SD, n = 3. **P < 0.01, vs. shCtrl. D Fractions of EPCAM-positive MDA-MB-231 cells were detected by flow cytometry. Mean ± SD, n = 3. **P < 0.01, vs. shCtrl. E Mammosphere formation of MDA-MB-231 cells was shown. Mean ± SD, n = 3. **P < 0.01, ***P < 0.001, vs. shCtrl. F Mammosphere formation of MCF7 cells was shown. Mean ± SD, n = 3. **P < 0.01, vs. OE Ctrl. G SOX2, CD133, and EPCAM expression in MCF7 cells were detected by real-time PCR. Mean ± SD, n = 3. ***P < 0.001, vs. OE Ctrl.

Fig 5: IGF2BP2 KO suppresses pancreatic cancer proliferation, which can be rescued by re-expression of IGF2BP2 in the KO cells.a KO of IGF2BP2 in BXPC-3 cells as verified by western blot. b IGF2BP2 KO suppresses cell proliferation in BXPC-3 cells as determined by MTT assay. c IGF2BP2 KO suppresses colony formation in BXPC-3 cells. d Re-expression of IGF2BP2 in IGF2BP2 KO cells as determined by western blot. e Rescue of IGF2BP2 promotes cell proliferation in two IGF2BP2 KO cell lines. f Rescue of IGF2BP2 increases colony formation in two IGF2BP2 KO cell lines. g IGF2BP2 KO suppresses stemness-related genes (OCT4, NANOG, CD24, and CD133), as determined by qRT-PCR. Values in b, c, e, f, g are mean ± SEM. **P < 0.01; ***P < 0.001.