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. 2020 Sep 18;8(2):e000517. doi: 10.1136/jitc-2019-000517

Figure 7.

Figure 7

MiR-99b attenuated M2 polarization via repressing the mTOR/IRF4 axis in macrophages. (A) Cell lysates from BMDMs transfected with simTOR or Ctrl were analyzed by western blot. The relative protein levels of mTOR, STAT3, p-STAT3 and IRF4 were quantitatively compared (n=3). (B) BMDMs were pretreated with DMSO or Torin1 for 6 hours, and then stimulated with PBS, LPS+IFNγ or IL-4. The IRF4 expression was examined by qRT-PCR (n=4). (C) BMDMs transfected with miR-99b mimics or Ctrl were stimulated with PBS, LPS+IFNγ or IL-4 for 24 hours. The relative protein levels of IRF4 were quantitatively compared by western blot (n=3). (D) BMDMs were transfected with miR-99b ASO or Ctrl and siIRF4 followed by PBS or IL-4 stimulation for 24 hours. The expression of indicated genes was determined by qRT-PCR (n=4). (E) Schematic diagram of TAM-targeted delivery of miR-99b inhibiting tumor growth by reprogramming TAM phenotype. Data are shown as mean±SEM. *p<0.05; **p<0.01; ***p<0.001 by unpaired student’s t-testonline supplementary additional file. BMDMs, bone marrow-derived macrophages.