Fig. 3

MiR-375 targets HOXB3 via binding 3′-UTR of HOXB3. a Schematic of putative binding sites for miR-375 in 3′-UTR of HOXB3. b 293 T cells were transfected with wide-type pMIR-HOXB3UTR (WT), pMIR-HOXB3UTR (Mut), pMIR-NC, and pRL-SV40 containing Renilla luciferase gene for 24 h, followed by the transfection with miR-375 mimic or Scramble for another 24 h. Firefly and Renilla luciferase activities were both detected and histograms showed that the Firefly luciferase activities were normalized to Renilla luciferase activities. c The expression of miR-375 was detected in HL-60 and THP1 cells transduced with MSCV-miR-375 or MSCV-NC. *P < 0.01 versus MSCV-NC. (d and e) HOXB3 protein and mRNA expressions were detected by western blot and qRT-PCR in HL-60 and THP1 cells following transduction with MSCV-miR-375 or MSCV-NC, respectively. *P < 0.01 versus MSCV-NC. f HOXB3 protein expression was measured in three primary AML blasts, which were transduced with MSCV-miR-375 or MSCV-NC. g HOXB3 protein expression was detected in HL-60 and THP1 cells transfected with special miR-375 inhibitor or Scramble for 48 h. (H) HOXB3 expressions were detected in 7 leukemic cell lines, 102 primary AML blasts, and 20 NC by qRT-PCR. i miR-375 and HOXB3 expressions were measured in 102 primary AML blasts and plotting of miR-375 expression versus HOXB3 expression showed an inverse correlation between them. The correlation coefficient (R) and P values were detected by Pearson correlation