HPDE6, PANC-1, BxPC-3, Capan-2, SW1990, and AsPC-1 cells lines were obtained from Procell (Wuhan, China). Cells were cultured in RPMI 1640 medium (Gibco, New York, USA), supplemented with 10% fetal bovine serum (FBS) at 37℃ with 5% CO2. PFT-α treatment was administered at a final concentration of 100 ng/ml for 24 h.
For the generation of cell lines with ZMAT1 overexpression or knockdown, the EGFP-tagged/3 × Flag hZMAT1-CMV Puro vector and three siRNAs targeting ZMAT1 were transfected into cells to overexpress and silence ZMAT1, respectively. Cell transfection was performed as previously described [17 (link)]. After antibiotic selection, the over-expression and depletion efficiency were assessed by Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Reconstituted cells with stable overexpression of ZMAT1 and cells with depleted ZMAT1 were utilized for cell proliferation assays, cell migration assays, RT-qPCR, immunoblotting, and animal experiments as indicated below. The siRNA and PCR primer oligonucleotide sequences used in our study are shown in Tables S1-S2.
For the generation of cell lines with ZMAT1 overexpression or knockdown, the EGFP-tagged/3 × Flag hZMAT1-CMV Puro vector and three siRNAs targeting ZMAT1 were transfected into cells to overexpress and silence ZMAT1, respectively. Cell transfection was performed as previously described [17 (link)]. After antibiotic selection, the over-expression and depletion efficiency were assessed by Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Reconstituted cells with stable overexpression of ZMAT1 and cells with depleted ZMAT1 were utilized for cell proliferation assays, cell migration assays, RT-qPCR, immunoblotting, and animal experiments as indicated below. The siRNA and PCR primer oligonucleotide sequences used in our study are shown in Tables S1-S2.