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. 2019 Sep 7;10(23):5671–5680. doi: 10.7150/jca.34972

Figure 5.

Figure 5

Transiently over-expressed miR-4433 inhibited the expression of Bcr-Abl and induced apoptosis in K562 cells. (A) Bioinformatics analysis of predicted miR-4433 binding sites on the 3'UTR of the ABL gene. A mutant ABL 3'UTR construct was prepared by mutating nucleotides in the seed region of the miR-4433 target site. (B) SAHA increased the expression level of miR-4433. K562 cells were exposed to SAHA (5 μM) for 48 hours and the miR-4433 expression level was determined by qRT-PCR. (C) K562 cells were transiently transfected with miR-4433, NC for negative control and C for control group, and then Bcr-Abl levels were determined by qRT-PCR. (D) Apoptosis-related proteins including PARP, caspase-3 were determined by western blot after the transfection in K562 cells. (E) p-Bcr-Abl, Bcr-Abl, p-Akt, Akt, p-Erk-1/2 and Erk-1/2 were examined by western blot post transfection in K562 cells. (F) Additionally, 48 hours post transfection of K562 cells, the apoptosis rates were measured by flow cytometry after Annexin V- FITC and propidium iodide (PI) double staining. Column, mean; bars, SE; **, p < 0.01, ***, p < 0.001, Student's t test.