Skip to main content
. 2019 Mar 29;9:5348. doi: 10.1038/s41598-019-41866-y

Figure 2.

Figure 2

Regulatory and functional interactions between the miR-221 network and P-gp in multidrug-resistant leukemia cells. (A–C) qPCR measuring the expression levels of (A) miR-221, (B) NCL, DNMT1 and (C) p15ink4b, p27kip1 in K562, DOX-treated K562 (10 µM, 24 h) and K562DR cells. Data represent three independent experiments and are expressed as the mean ± SD; **P < 0.01 (D) Western blot showing NCL, DNMT1 and p27kip1 protein expression in K562, DOX-treated K562 (10 µM, 24 h) and K562DR cells. (E) Immunofluorescence assay showing P-gp expression in K562 and K562DR cells (blue, hoechst; green, P-gp; light, giemsa staining of leukemia cells). (F) Western blot showing P-gp protein expression in K562, DOX-treated K562 (10 µM, 24 h) and K562DR cells. (G) Immunofluorescence assay showing P-gp expression in K562 cells transfected with miR-221 or a scrambled control and in K562DR cells transfected with anti-221 or a scrambled control (blue, hoechst; green, P-gp). (H) Western blot showing p27kip1 and P-gp protein expression in K562 and K562DR cells transfected with miR-221, anti-221 or a scrambled control. (I) Spearman correlation analysis of miR-221 and P-gp RNA levels in AML patients (109 AML patients, GEO database GSE68466 and GSE68467). (J) K562 (above) and K562DR (below) cells were transfected with miR-221, anti-221 or scrambled control for 6 hours, treated with doxorubicin (0 to 10 µM) for an additional 72 hours and cellular viability was measured with the CCK-8 assay. (K) Summary diagram describes regulatory and functional interactions between the miR-221 network and P-gp in multidrug-resistant leukemia cells.