Fig. 6: Validation of MIR1307-mediated effect in an independent in vitro PDAC model.

A Capan 1 cells were stably infected with miRZip lentiviral vectors expressing MIR1307-sh or CTRL probes. MIR1307. A miRZip MIR1307-sh cells had reduced MIR1307 expression when detected by Taqman assay. Please note that it is recognized that Taqman assays can detect the MIR1307-sh produced by the vector and therefore justifies the lack of >90% MIR1307 expression. Bars represent mean of three biological replicates with SD. Values from two-sided t-test are reported. B CLIC5mRNA was assessed by real time PCR. Bars represent the mean and SD of three biological replicates. Values from two-sided t-test are reported. C CLIC5 and vinculin protein expression were assessed by western blotting. D Cells were plated in 96-well plates and treated with DMSO or FOI for 48 h before assessing cell viability by CellTiter Blue. Bars represent the mean and SD of 12 replicates repeated in two separate occasions. Values from two-sided t-test are reported. E Capan 1 cells were treated with scalar concentrations of FOI (from 0.5 to 10 μM) in presence of absence of caspase 9 inhibitor for 48 h before being assessed for caspase activation. Bars represent the mean and SD of three independent replicates. F Capan 1 cells were treated for 48 h and COMET assay performed afterwards. Representative images are shown (magnification/scale 20×, magnification bar indicates 50 μm). At least 50 cells (across three replicates) were assessed for each sample. Bars represent mean and standard error. Values from two-sided t-test are reported. G Cells were treated with DMSO or FOI for 24 h and fixed before being stained with the indicated fluorescent antibodies. Representative pictures (left) along with quantitation (right) of the percentage of positive cells on the total cells in the field, with at least five fields assessed per replicate in six biological replicates. Bars represent the mean and 95% Confidence interval. The magnification bar indicates 20 μM.