Fig. 5: MIR1307-dependent effects are mediated by modulation of CLIC5 expression.

A mRNA expression of indicated genes was assessed by TaqMan assay. Bars indicated the mean and SD of three replicates. Values from two-sided t-test are reported. B Cells were transfected for 48 h and treated with FOI for additional 48 h before mRNA expression was assessed by TaqMan assay. Bars represent mean and SD of six replicates normalized on the WT siCTRL mimicCTRL sample. Values from two-sided t-test are reported. C Cells were transfected for 48 h and treated with FOI for 48 h and then collected for protein assessment by western blotting. Representative pictures and quantification plots are shown here. Values from two-sided t-test are reported. Source data are provided as a Source Data file. D Luciferase assay was performed in MiaPaca2 cells using the vectors indicated. MIR1307 or scrambled mimic were added as indicated. Data are presented normalized to the second column from the left indicating cells transfected with CLIC5 WT vector and scrambled mimic. Bars represent the mean and SD of three independent replicates. Values from two-sided t-test are reported. E Cells were transfected for 48 h and treated with FOI for 48 h before assessed for cell viability by CellTiter-Blue assay. Bars represent the mean and SD of six replicates. Values from two-sided t-test are reported. Bars below the 0 line indicate reduced cell viability in FOI sample vs DMSO sample. F MIR1307KO cells were transfected with the indicated siRNA and DMSO or FOI chemotherapy added 24 h later. 8OHdG (marker of DNA damage from oxidative stress) was measured after 48 h of treatment. Bars indicate mean and SD of six replicates. Values from two-sided t-test are reported. As the value generated by the assay is inversely related to the DNA damage, values here are expressed as 1/actual data. G MIR1307KO cells were transfected with the indicated siRNA and DMSO or FOI chemotherapy added 24 h later. COMET assay was performed after 48 h of treatment. At least 200 measurements from more than five biological replicates were taken for each condition. Bars represent mean and SE. Values from two-sided t-test are reported. H Cells were treated with DMSO or FOI for 24 h in the presence or absence of 10 mM N-acetyl-cystein (a ROS scavenger) and ROS detected and quantitated by a luminescence-based assay. Menadione 50 μM was used as positive control (inducer of ROS). Bars represent the mean and SD of three biological replicates. * indicate p < 0.05 from two-sided t-test are reported.