Fig. 5

miR-4532 upregulation represses hematopoiesis of HSCs by activating LDOC1-dependent STAT3 signaling pathway. a The extents of JAK2 and STAT3 phosphorylation in CD34⁺ HSCs treated with different dose of exosomes, as determined by Western blot analysis (*p < 0.05 vs. CD34⁺ HSCs treated with PBS). b The extents of JAK2 and STAT3 phosphorylation in CD34⁺ HSC culture with transfected Molm-14 cell-derived exosomes, as determined by Western blot analysis (*p < 0.05 vs. CD34⁺ HSCs co-cultured with exosomes from inhibitor-NC-treated Molm-14 cells). c The extents of JAK2 and STAT3 phosphorylation in CD34⁺ HSCs after alteration of miR-4532 and LDOC1 determined by Western blot analysis (*p < 0.05 vs. CD34⁺ HSCs treated with mimic-NC, ^#p < 0.05 vs. CD34⁺ HSCs cultured with miR-4532 mimic, ^&p < 0.05 vs. CD34⁺ HSCs cultured with si-NC). d The extents of JAK2 and STAT3 phosphorylation in CD34⁺ HSCs after inhibition of JAK2 determined by Western blot analysis. e Colony-forming capacity of CD34⁺ HSCs after inhibition of JAK2. f The relative expression of hematopoietic suppressor gene DKK1 in CD34⁺ HSCs after inhibition of JAK2 measured by RT-qPCR. *p < 0.05 vs. CD34⁺ HSCs treated with DMSO. Measurement data were described as mean ± standard deviation. Comparison between two groups was 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; LDOC1, leucine-zipper downregulated in cancer 1; RT-qPCR, reverse transcription quantitative polymerase chain reaction; STAT3, signal transducer and activator of transcription 3