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. 2020 Dec 3;25(1):575–585. doi: 10.1111/jcmm.16110

Figure 3.

Figure 3

miR‐15a‐5p and miR‐21‐5p regulate the expression of ARL2, PDCD4 and BTG2. A, Quantitative RT‐PCR of PDCD4, ARL2 and BTG2 in K562 after 24 h of transfection with synthetic miR‐15a‐5p, miR‐21‐5p or SCR treated with cytarabine and daunorubicin. The results are shown as average mRNA expression after normalization with GAPDH and 2ΔCt calculations. B, Western blotting of PDCD4 and ARL2 protein expression in K562 cells treated with cytarabine and daunorubicin after 24 h of transfection with synthetic miR‐15a‐5p, miR‐21‐5p or SCR. The protein loading control was performed using β‐actin. C, Bands were quantified by densitometry using Image J software. Data represent the average of three independent experiments ± SD. P values were obtained using a t test. * Indicates a significant difference P ≤ .05, and ** indicates a significant difference P ≤ .01. D, Luciferase activity in HEK293 cells cotransfected with synthetic miR‐15a‐5p, miR‐21‐5p or SCR and luciferase reporter constructs containing wild‐type (WT) or mutated (miRNA_Δ) PDCD4, ARL2 and BTG2 3′UTR. Luciferase activities were determined at 24 h and were normalized using β‐galactosidase activity. The mutant plasmids were generated by deleting the miR‐15a‐5p or miR‐21‐5p binding site(s)