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. 2016 Aug 2;7(50):82338–82353. doi: 10.18632/oncotarget.11001

Figure 5. MiR-891b targeted the regulation of the downstream signal molecules of Cbl-b.

Figure 5

(A) MiR-891b was overexpressed, or Cbl-b and Smad3 were silenced in PDAC cell lines, SW1990 and PANC-1. The expressions of associated proteins were detected by western blot. (B) The PDAC cells were transfected with miR-891b mimic/NC, or Cbl-b siRNA/NC for 48h, and cell culture supernatants were collected. Active TGF-β1 levels were determined by Elisa. *P < 0.05. (C–D) After pretreated with or without 10 μmol/L SB431542 for 2 h, PDAC cells were transfected with miR-891b mimic/NC, or Cbl-b siRNA/NC. (C) Cells were collected at 48 h after the transfection. The cell proliferation was determined by counting the non-trypan blue staining cells. *P < 0.05. (D) The expression of Cbl-b, Smad3, p-Smad3 and p21 were detected by western blot analysis. (E) After miR-891b mimic/NC or Cbl-b siRNA/NC was transfected in PANC-1 cell line, subcellular localization of Smad3 was identified using immunofluorescence. DAPI, 4′-6-Diamidino-2-phenylindole. (F) The PDAC cells were transfected with miR-891b mimic/NC, or Cbl-b siRNA/NC. The Smad3 mRNA level was determined by qRT-PCR. 18S ribosomal RNA was used as an internal control for relative quantitation. *P < 0.05.