Fig. 2

Specific interaction between miR-125a-5p and St3gal1 was verified by RNAi and dual luciferase reporter assay. A Transfection efficiency of the murine intestinal cell line CMT 93 with miR-125a-5p mimics as well as expression of potential targets was evaluated by means of RT-qPCR. Most pronounced and significant decrease of the target St3gal1 was detected after miR-125a-5p transfection. B4galt1 showed significantly reduced levels. St3gal2 showed no significant difference. miR-615-3p showed upregulated but not significant expression. Non-target miRNA was used as a control, fold changes were calculated relatively to the non-target control and normalised with HPRT and SDHA or SNORD44 and SNORD 47, respectively. Charts indicate means ± standard deviations (SD) of three biological samples with triple measurements. B Relative luciferase activity was determined in comparison to control non-target miRNA mimic. miR-125a-5p caused clear and significant activity of the reporter gene fused to the target sites compared with non-target transfected controls. Charts indicate means ± SD of three biological replicates with technical triplicates. Statistical significance is presented by asterisks compared to negative controls at each time point. *P ≤ 0.05, **P ≤ 0.01, unpaired t-test