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. 2015 Aug 18;10(8):e0135867. doi: 10.1371/journal.pone.0135867

Fig 4. MiR-200c regulates the expression of FOG2 and PTEN.

Fig 4

(A) Effect of miR-200c on the luciferase activity. 3’-UTR conserved site of PTEN and its mutated version for miR-200c pairing were exhibited in upper part. 293T cells were cotransfected with the luciferase reporter vectors and miR-200c mimics (miR-200cM) or control oligoes (Ctr.oligoes). The luciferase activity was measured after transfection for 36 hrs according to the protocol described in Materials and Methods (lower). Vector, pSiCHECK-2 vector; WT 3’-UTR, wild type PTEN 3’UTR; Mut-3’-UTR, PTEN 3’UTR with mutation in the putative miR-200c binding sites. (B) Effects of MiR-200c on the expression of PTEN and FOG2. MDSCs were transfected with miR-200c mimics (miR-200cM), miR-200c inhibitors (miR-200cI) or control oligoes (Ctr. oligoes),and then protein of PTEN and FOG2were detected by Western blot. The right histogram represents the relative densitometric value corresponding bands in the Western blot. (C) Effects of TCCM on the expression of miR-200c, PTEN and FOG2. BM cells were exposed to TCCM and then the expression of miR-200c (upper) and PTEN (middle) and FOG2 (lower) was detected at the indicated time points. BMC, bone marrow cells which were not exposed to TCCM. (D) Effect of anti-miR-200c on the expression of PTEN and FOG2 in TCCM treated BM cells. BM cells were transfected by miR-200c inhibitors (miR-200cI) or control oligoes (Ctr. oligoes), and then exposed to TCCM. Transcriptional levels of PTEN (upper) and FOG2 (lower) were detected by qRT-PCR at the indicated time. (E) Protein levels of PTEN and FOG2 in BM cells after exposed to TCCM. Protein levels of PTEN and FOG2 in BM cells after exposed to TCCM were detected by Western blot at the indicated time. Each experiment was independently performed three times. *, p<0.05; **, P<0.01.