Fig. 5: Functional impact of CFTR overexpression and CFTR mutants on PANC-1.

a, b PANC-1 cells transduced with lentivirus carrying wild-type CFTR or CFTR mutants were cultured for 2 weeks. CFTR RNA expression levels (a) were examined using qPCR (n = 3 technical replicates/group). CFTR protein and FLAG tags levels (b) were examined using Western blot. c–e The proliferation of PANC-1 cells were assessed using CCK-8 method (c, n = 5 technical replicates/group), colony formation assay (d, n = 3 technical replicates/group) and EdU incorporation assays (e, n = 6 technical replicates/group). f The migration and invasion of PANC-1 cells were measured following CFTR overexpression using the Transwell assay (Magnification: ×200, scale bar = 200 μm) (n = 9 technical replicates/group). g The apoptosis of PANC-1 cells using Caspase 3/7 assay (n = 3 technical replicates/group). h Tumor growth was measured at day 22 of inoculation (n = 5 biological replicates/group). In the accompanying photograph of mice bearing xenograft tumors, the tumors’ locations are indicated by arrows. Statistical significance was assessed by two-sided t-test. i The immunofluorescence staining of CFTR in PANC-1. Magnification: ×630, scale bar = 20 μm. Five times each experiment was repeated independently with similar results. j Western blot of the cell fractionation experiments. CFTR were detected in membrane fractions in PANC-1 CFTR OE, p.R31C, p.E681V and p.C1355Y. Three times each experiment was repeated independently with similar results. k Gene ontology analysis by comparing the EV and CFTR OE conditions. l Heatmap visualizations of pathway based on ssGSEA analysis for EV, p.L88X, p.C1355Y, and OE, showing comparisons between each mutant and the CFTR OE. Statistical analysis was performed using one-way ANOVA other than (h); multiple comparisons were performed according to Tukey’s method. Data were presented as mean ± standard deviation (SD). Bars represent the SD with minima (lower bar) and maxima (upper bar) indicated. P-values for comparisons with EV are indicated by asterisks, with OE are indicated by pound signs. ns: not significant; */#P < 0.05; **/##P < 0.01; ***/###P < 0.001; ****/####P < 0.0001. Statistical significance among PANC-1 cells comparisons is tabulated in Supplementary Table 2. Source data are available as a Source Data file.