Figure 4.

ASPP2 is required for pancreatic cancer cell proliferation. (A,B) Establishment of ASPP2-inducible knockdown PANC-1 (A) and S2.013 (B) cell lines with a TetOn inducible system. (C,D) Cell proliferation assays in PANC-1 (C) and S2.013 (D) cells from A and B. ***: p = 0.00054 (shRNA-A vs. control) and p = 0.0085 (shRNA-B vs. control) in panel C. ***: p = 6.99 × 10⁻⁵ (shRNA-A vs. control) and 0.00019 (shRNA-B vs. control) in panel D. Data are expressed as the mean ± SD of three independent experiments. (E–H) Tumor growth curve (E), representative images (F), end-point tumor weight (G), and Ki-67 staining (H) of ASPP2-depleted S2.013 cells as well as the control counterparts in xenograft mouse models. Cells were subcutaneously inoculated into athymic nude mice (seven mice in each group). Doxycycline (1 mg/mL) was added to the diet on day six. Data are expressed as the mean ± SEM. *: p < 0.05; **: p < 0.01. (I,J) Establishment of ASPP2-inducible overexpression Capan-2 (I) and BxPC-3 (J) cell lines with a TetOn inducible system. (K,L) Cell proliferation assays in Capan-2 (K) and BxPC-3 (L) cells from I and J. Statistical significance between the ASPP2-overexpressed (ASPP2-WT and ASPP2-2A, 2A: S562A/S704A) cells is indicated by p values in the panels. Data are expressed as the mean ± SD of three independent experiments. Student’s t test was used to analyze the statistical significance of this figure. The uncropped bolts are shown in Supplementary Materials.