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. 2024 Sep 3;5(9):101711. doi: 10.1016/j.xcrm.2024.101711

Figure 3.

Figure 3

The upregulation and function of p63 in PPSSC cancer cells with adenosquamous phenotype

(A) Detection of p63 and CK5 in KPPC and PPSSC cancer cell lines by western blot assay.

(B) Quantitative reverse-transcription PCR (qRT-PCR) analysis of TAp63 and ΔNp63 expression in KPPC and PPSSC cancer cells (n = 3 biological replicates). Gene expression levels were compared with Student’s t test.

(C) Cell viability of KPPC and PPSSC cancer cell lines (n = 3 biological replicates) after transfection with siRNA-Trp63 (si-Trp63) or non-targeting siRNA-control (si-Ctrl). Cell viability was presented as a percentage normalized to the si-Ctrl group and compared with Student’s t test.

(D) Detection of p63 by western blot assay in KPPC and PPSSC cancer cell lines transfected with indicated siRNAs.

(E and F) qRT-PCR analysis of Notch1/Notch2 (E) or Irf3/Irf7/Ifit1 (F) expression in KPPC and PPSSC cancer cells (n = 3 biological replicates). Gene expression levels were compared with one-way ANOVA with Tukey’s multiple comparison test.

(G) GSEA results of RNA-seq data on KPPC and PPSSC cancer cell lines. Interferon-α and interferon-γ pathway genes were enriched in PPSSC cancer cells, as compared with KPPC cancer cells.

(H) qRT-PCR analysis of Cebpa, Cebpb, Cebpg, and Cebpd expression in KPPC and PPSSC cancer cells (n = 3 biological replicates). Gene expression levels were compared with one-way ANOVA with Tukey’s multiple comparison test.

(I) Schematic of syngeneic orthotopic pancreatic tumor formation experiments in C57BL/6J mice using KPPC and PPSSC cancer cells. Tumor-bearing mice were sacrificed for further analysis at 2 months after orthotopic implantation of cancer cells.

(J) Representative images of H&E staining and p63 immunohistochemistry staining on KPPC and PPSSC orthotopic pancreatic tumor sections (n = 5/group). KPPC pancreatic tumors were collected 1 month after orthotopic injection, while PPSSC pancreatic tumors were collected 2 months after orthotopic injection. Scale bar: 100 μm. ∗p < 0.05, ∗∗p < 0.01. See also Figures S3B and S3C.