Fig 1: The interaction of GOLPH3 and STIP1 regulates telomerase activity in PDAC cells. (A) GOLPH3 siRNA downregulates GOLPH3 expression mRNA levels in PANC1 and BXPC3 cells. (B) GOLPH3 siRNA downregulates GOLPH3 but not STIP1 levels in PANC1 and BXPC3 cells. (C) Expression of GOLPH3 and hTERT transfected with control siRNA and GOLPH3 siRNA-1/2 were assessed using qRT-PCR. (D) The relative activity of telomerase in the control and GOLPH3 knockdown of PDAC cells detected using q-TRAP. (E,F) Expression of c-Myc and Cyclin D1 transfected with the control siRNA and GOLPH3 siRNA-1/2 were assessed using qRT-PCR and western blotting (E mRNA and F Protein). (G) Expression of Cyclin D1, c-Myc, and GOLPH3 transfected with the control siRNA and STIP1 siRNA-1/2, assessed using qRT-PCR. (H) The relative activity of telomerase in the control and STIP1 knockdown PDAC cells detected using q-TRAP. *p < 0.05 and **p < 0.01.
Fig 2: GOLPH3 correlates positively with STIP1 in PDAC tissues. (A) Immunohistochemical staining for GOLPH3 and STIP1 in PDAC tissues (T) and paired normal pancreatic tissues (ANT). (B) Co-expression of GOLPH3 and STIP1 in PDAC tissues as assessed using immunofluorescence. (C) Expression of GOLPH3 analyzed using qRT-PCR in paired normal and PDAC tissues. (D) STIP1 levels were measured in paired PDAC tissues and adjacent normal tissues using qRT-PCR. The data were normalized to GAPDH levels as a control. (E) Correlation of GOLPH3 and STIP1 expression in PDAC tissues. (F) GOLPH3 and STIP1 protein levels were measured in paired PDAC tissues using western blotting. (G) The expression of GOLPH3 and STIP1 in different PDAC cell lines compared with that in pancreatic cyst tissue as a control.
Fig 3: Knockdown of STIP1 inhibits cell proliferation. (A) STIP1 siRNA inhibited proliferation of PDAC cells as detected using CCK-8 assays at different time points. (B) STIP1 siRNA inhibited colony formation in pancreatic cancer cells. (C) Cell cycle assay to determine the role of STIP1 in cell cycle progression after downregulating STIP1. (D) EdU staining assay to determine the effect of STIP1 downregulation on cell proliferation. *p < 0.05, **p < 0.01, and ***p < 0.001.
Fig 4: GOLPH3 promotes the tumorigenicity of PDAC cells in vivo. (A) Xenograft model in nude mice; representative image of tumors from all mice in each group. (B) Dots graphs represent the weight of the BXPC3 cells xenograft tumor after sacrificing mice. (C) Tumor volume growth curves by the indicated cells. Growth of subcutaneous xenografts was weakened by downregulation of GOLPH3 in nude mice. (D) Time courses of animal weight. (E) IF assay to detect GOLPH3 and STIP1 localization in subcutaneous xenografts. (F) HE staining and IHC of GOLPH3, STIP1, ki67, Cyclin D1, and Bim in subcutaneous xenografts in nude mice. (G) qRT-PCR analysis of GOLPH3, c-Myc, and hTERT mRNA expression in vivo. (H) The relative activity of telomerase in paired PDAC tissues in the xenograft tumors (upper panel) and GOLPH3, c-Myc, and hTERT expression by western blotting analysis (lower panel). (I) The relative expression of GOLPH3 in GOLPH3 shRNA group and vector group. *p < 0.05, **p < 0.01, and ***p < 0.001.
Fig 5: GOLPH3 interacts with STIP1. (A) A schematic diagram of the BiFC screening assay. Non-fluorescent subfragments of a fluorescent protein YFPn and YFPc were fused to X and Y, respectively. If X and Y interact, the two fragments could associate and refold, allowing fluorescence to occur. (B) BiFC screening for GOLPH3 interacting proteins and a map of the BiFC results for protein interaction networks constructed using STRING. (C) FACS analysis in HTC75 cells co-expressing BiFC variants of GOLPH3, VPS35, and STIP1; and quantitative analysis of YFP fluorescence intensity from the panel represented as a histogram. (D) The Co-IP result showing the interaction between GOLPH3 and VPS35 and STIP1. (E) Immunofluorescence showing GOLPH3 and STIP1 co-expressed in PDAC cells.
Supplier Page from Abcam for Anti-STIP1/STI1 antibody [EPR6606]