Figure 4: CASP9 is directly targeted by miR-224. (a) CASP9 transcript expression in MDA-MB-231 cells was measured 48 hours posttransfection with antimiR-224 (40 μM) or negative control miR (40 μM). (b) Western blotting of CASP9 in MDA-MB-231 cells was performed 72 hours posttransfection. (c) CASP9 expression was measured in TNBC samples. (d) Schema shows miR-224 interaction with its target mRNAs—CASP9 and BRAF1. miRNA seed sequence was shown in red. (e) Luciferase reporter assay of MDA-MB-231 cells cotransfected with pCASP9-UTR or pCASP9 Mutant plasmids and either antimiR-224 or negative control miR. Samples were analyzed 72 hours posttransfection, and data were normalized to the pmiR plasmid only transfection. Data represents the ; and . (f) Caspase-3/7 activities were measured at 48 and 72 hours posttransfection with untreated, antimiR control, antimiR-224 (40 μM/well), or antimiR-224 (80 μM/well). Each datum represents the from 3 independent experiments.