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Fig. 1 | Stem Cell Research & Therapy

Fig. 1

From: Acute myeloid leukemia cells secrete microRNA-4532-containing exosomes to mediate normal hematopoiesis in hematopoietic stem cells by activating the LDOC1-dependent STAT3 signaling pathway

Fig. 1

miR-4532 expression is upregulated in AML cell-secreted exosomes. a Screening of AML-related miRs in the GSE85769 microarray data. The X axis represents the sample number, and the Y axis represents the gene name. The histogram on the upper right is color gradation, with each rectangle representing a corresponding sample expression value. b miR-4532 expression in AML cell lines (HL-60, Molm-14, ML-2, and OCI-AML3) and CD34+ HSCs, as measured by RT-qPCR. *p < 0.05 vs. CD34+ HSCs. c Schematic diagram of exosome separation procedure. To exclude the effect of exosomes on bovine, the medium or FBS was centrifuged at 100,000×g for 10 h to remove the bovine exosomes. After that, the centrifugal medium was filtered through a 0.2-μm filter and collected for cell culture. AML cell line was cultured with the centrifugal medium. After 48 h, the supernatant was obtained. The process of exosome separation is shown in c. Finally, the purified exosomes were rinsed twice with PBS. d Morphology of exosomes observed under TEM. e The size and concentration of exosomes evaluated by nanoparticle tracking analysis. f Identification of marker exosomes (TSG101, CD63, and histone) by Western blot analysis. g miR-4532 expression in AML cell lines (HL-60, Molm-14, ML-2, and OCI-AML3) and exosomes secreted from AML cell lines (HL-60, Molm-14, ML-2, and OCI-AML3), as measured by RT-qPCR. *p < 0.05 vs. HL-60 cells, Molm-14 cells, ML-2 cells, or OCI-AML3 cells. Measurement data were described as mean ± standard deviation. Comparisons between two groups were analyzed by unpaired t test, and comparisons among multiple groups were analyzed by one-way analysis of variance (ANOVA), followed by Tukey’s post hoc test. The cell experiment was repeated three times to obtain the mean value. miR-4532, microRNA-4532; RT-qPCR, reverse transcription quantitative polymerase chain reaction; TEM, transmission electron microscope; PBS, phosphate buffer saline; FBS, fetal bovine serum

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