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. 2017 Nov 18;8(63):107188–107205. doi: 10.18632/oncotarget.22513

Figure 3. The NRF2-silencing-induced miR-206 directly mediates c-MET and EGFR reductions.

Figure 3

(A-B) SKOV3 (A) or A498 (B) cells were co-transfected with miR-206 and the c-MET-3′-UTR- or EGFR-3′UTR-luciferase reporter plasmid. After 18 h, 3-UTR-derived luciferase activity was measured. Values are means ± SD from 4 experiments. (C-D) After transfection of shNRF2-SKOV3 (C) or shNRF2-A498 (D) cells with the miR-206 inhibitor (100 nM) or negative control (NC), c-MET and EGFR protein levels were assessed by Western blotting. Data are means ± SD from three independent experiments. (E) The scSKOV3 cells were incubated with trichostatin (TSA, 0.3 μM) for 18 h, and miR-206 levels were measured using real-time PCR analysis. Values are means ± SD from three experiments. (F-G) Protein levels for c-MET and EGFR were determined in SKOV3 (I) and A498 (J) following the incubation with TSA (0.3 μM) for 24 h. (H) The transcript level of HDAC2 was assessed in sc and shNRF2-SKOV3 cells. (I) Cytoplasmic (Cyto) and nuclear (Nuc) levels of HDAC2 were determined Western blotting. (J) The scSKOV3 cells were transiently transfected with nonspecific siRNA (siCTL) or HDAC2-specific siRNA (siHDAC2) and the protein levels for c-MET and EGFR were assessed by Western blotting. Similar blots were obtained from three independent experiments (D, F, G, I and J).