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. 2020 Aug 3;20(4):3661–3668. doi: 10.3892/etm.2020.9088

Figure 3.

Figure 3

Inhibition of ALK5 aggravates high-dose TGF-β1-induced degeneration of NPCs. NPCs were treated with 5 nM TGF-β1 or 100 nM SB for 72 h. For the co-treated group, NPCs were treated with 5 nM TGF-β1 combined with 100 nM SB for 72 h. (A) Immunofluorescence staining of TGF-β1 (magnification, x400) and (B) quantification analysis. (C) Protein expression of ALK5 and ALK1 was determined by western blotting and (D) semi-quantified. (E and F) The mRNA expression levels of smad1/2/3/5/8, Col II, TIMP-3, Col I and MMP-13 in NPCs were determined using reverse transcription-quantitative PCR. Data are presented as the mean ± SD of three independent experiments. *P<0.05, **P<0.01, ***P<0.001. TGF-β1, transforming growth factor β1; NPC, nucleus pulposus cell; ALK, ALK tyrosine kinase receptor; Col, collagen; TIMP-3, tissue inhibitor of metalloproteinase-3; MMP-13, matrix metalloproteinase 13; SB, SB525334.