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. 2019 Jan 1;10(1):45–59. doi: 10.18632/oncotarget.26534

Figure 4. Validation of miRNA and their potential targets.

Figure 4

(A) SHSY 5Y cells were cultured with either DMSO or 10 µM CBD as described in Figure 1 legend. Next, qPCR was performed for the genes of interest according to Ingenuity Pathway Analysis map. (B) SH SY5Y cells were treated with either mock, hsa-let-7a-mimic or hsa-let-7a inhibitor for 24 hours. Next, qPCR was performed. We validated that hsa-let-7a transfection was successful as shown in the first panel. The next two panels show the levels of expression of caspase-3 and GAS7. (C) SH SY5Y cells were treated with mock, hsa-miRNA-1972 mimic or hsa-miRNA-1972 inhibitor. Next, qPCR was performed. First panel shows validation that hsa-miRNA-1972 transfection was successful. The next three panels show the levels of expression of BCL2L1, Sirt-2, and MYCN. (D) After transfecting SH SY5Y with hsa-let-7a mimic and inhibitor, protein was obtained and western blot was done to detect the level of caspase-3. The level of caspase-3 protein was 4.7 folds higher than the mock control. Significance (p < 0.05) for Panel A was performed by using Student’s t-test and for Panel B and C was determined using one-way ANOVA and post-hoc Tukey’s test.