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. 2019 Feb 11;2019:7378967. doi: 10.1155/2019/7378967

Figure 4.

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 mean ± SE; n = 3 and ∗ p < 0.05. (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 mean ± SE from 3 independent experiments.