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. 2021 Aug 8;12:2040622321997259. doi: 10.1177/2040622321997259

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© The Author(s), 2021

This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 4. — miR-328-3p inhibits AML cells resistant to Ara-C by targeting CCND2. (a) CCND2 expression in AML according to TCGA (bone marrow tissue samples from 173 AML patients) and GTEx (70 normal bone marrow samples) databases, *p < 0.05 compared with normal group. (b) Venn diagram to show the miRNA targets of MALAT1 and CCND2 according to the analysis from starBase. (c) RT-qPCR to detect the expression of MALAT1 in bone marrow tissue samples from AML patients and normal bone marrow samples, *p < 0.05 compared with normal group. (d) Dual-luciferase reporter assay to detect the relationship between miR-328-3p and CCND2, *p < 0.05 compared with NC mimi. (e) RT-qPCR to detect the expression of CCND2 after overexpression of miR-328-3p in HL-60/A cells, *p < 0.05 compared with NC mimic. (f) RT-qPCR to detect the expression of miR-328-3p and CCND2 after overexpression of miR-328-3p and CCND2 in HL-60/A cells, *p < 0.05 compared with NC mimic + oe-NC, ^#p < 0.05 compared with miR-328-3p mimic + oe-NC. (g) Western blot of the expression of CCND2 after overexpression of miR-328-3p and CCND2 in HL-60/A cells, *p < 0.05 compared with NC mimic + oe-NC, ^#p < 0.05 compared with miR-328-3p mimic + oe-NC. (h) MTT assay to detect the viability of HL-60/A cells after overexpression of miR-328-3p and CCND2, *p < 0.05 compared with HL-60/miR-328-3p mimic + oe-NC, ^#p < 0.05 compared with NC mimic + oe-NC/miR-328-3p mimic + oe-CCND2. (i) Flow cytometry to analyze the cell cycle of HL-60/A after overexpression of miR-328-3p and CCND2, *p < 0.05 compared with NC mimic + oe-NC, ^#p < 0.05 compared with NC mimic + oe-CCND2. Experiments were repeated three times. Data from two groups were compared by unpaired t-test. Data among multiple groups were analyzed by one-way analysis of variance.

AML, acute myeloid leukemia; Ara-C, cytarabine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Mut, mutant; NC, negative control; oe, overexpression; RT-qPCR, real-time quantitative polymerase chain reaction; TCGA, The Cancer Genome Atlas; Wt, wild type; GTEx, Genotype-Tissue Expression; miRNA, microRNA.